Composition based on substituted 1,3-diphenylprop-2-en-1-one derivatives, preparation and uses thereof

ABSTRACT

The invention concerns compositions comprising substituted 1,3-diphenylprop-2-en-1-one derivatives designed for therapeutic use. The inventive compositions are useful in particular for preventing or treating cardiovascular diseases, syndrome X, Ia restenosis, diabetes, obesity, hypertension, inflammatory diseases, cancers or neoplasms (benign or malignant tumors), neurodegenerative, dermatological diseases and disorders related to oxidative stress, for preventing or treating the effects of ageing in general and for example skin ageing, in particular in the field of cosmetics (occurrence of wrinkles and the like).

This application is a divisional of application Ser. No. 12/609,270(allowed), filed Oct. 30, 2009 (published as US-2010-0120909-A1), whichis a continuation of application Ser. No. 10/520,078 (issued as U.S.Pat. No. 7,632,870 on Dec. 15, 2009), filed Apr. 4, 2005 (published asUS-2005-0171149-A1), which is a U.S. national phase of InternationalApplication PCT/FR2003/002128, filed Jul. 8, 2003, which designated theU.S. and claims benefit of FR 02/08570, filed Jul. 8, 2002, the entirecontents of each of which is incorporated herein by reference.

The invention concerns compositions comprising substituted1,3-diphenylprop-2-en-1-one derivatives and uses thereof, particularlyin the fields of human and veterinary health. The compounds according tothe invention have pharmacological antioxidant properties and PPARα andPPARγ activator properties which are advantageous. The invention morespecifically describes the different uses of said compounds and thepharmaceutical and cosmetic compositions comprising same. The compoundsaccording to the invention are useful in particular for preventing ortreating cardiovascular diseases, syndrome X, restenosis, diabetes,obesity, hypertension, inflammatory diseases, cancers or neoplasms(benign or malignant tumors), neurodegenerative, dermatological diseasesand disorders related to oxidative stress, for preventing or treatingthe effects of ageing in general and for example skin ageing, inparticular in the field of cosmetics (development of wrinkles and thelike).

The derivatives and/or compositions of the invention can be used inparticular for treating diseases implicating tissues that express PPARαand PPARγ. More particularly, they advantageously allow to treatinflammatory, proliferative, degenerative diseases or pathologiesaffecting different organs and tissues, in particular diseases involvingpathological angiogenesis or neovascularization as well as any pathologyor disorder (for example related to age) involving an oxidative stress.In an advantageous manner, the compounds of the invention can be used inthe treatment of diseases or disorders affecting tissues and/or organs,irrespective of the etiological component.

The PPARs, or “peroxisome proliferator activated receptors”, are nuclearreceptors from the superfamily of transcription factors activated by thefollowing ligands: steroids/thyroid hormones/retinoids. To date, threePPAR isotypes have been cloned in mice and humans: PPARα, PPARβ/δ andPPARγ. While PPARβ/δ expression in humans appears to be ubiquitous,PPARα and γ exhibit a differential tissue distribution (Braissant andWahli 1998). PPARα is expressed in cells with high fatty acid catabolicactivity and in cells with high peroxisomal activity (hepatocytes,cardiomyocytes, renal proximal tubules, intestinal mucosa). PPARβ/δ isexpressed ubiquitously and abundantly in most tissues. As far as PPARγexpression is concerned, it is limited mainly to adipose tissue, certainimmune system cells and retina and is present in only trace amounts inother organs (Braissant and Wahli 1998).

The PPARs contain several domains having different properties. A DNAbinding domain (DBD) recognizes specific sequences, also called responseelements, located in regulatory regions of their target genes. Likeother nuclear receptors, the PPARs also contain a ligand binding domain,the activation of PPARs by their ligand modulating the expression ofgenes which contain specific PPAR response elements (PPRE) in thepromoter region. To activate transcription of their target genes, theactivated PPARs must heterodimerize with another nuclear receptor, RXR(Retinoid-X-Receptor). Taking the example of PPARα, its action ismediated by a class of compounds such as the fibrates which have alipid-lowering effect. Natural ligands have also been identified such asfor example fatty acids, eicosanoids (leukotriene B₄) and8(S)-hydroxyeicosatetraenoic acid (Kliewer, Sundseth et al. 1997).

The PPARs have been associated primarily with lipid and glucosemetabolism. PPAR activators, such as fibrates, enable a regulation ofplasma cholesterol and triglyceride concentrations via activation ofPPARα (Hourton, Delerive et al. 2001). Fibrate therapy leads to anincrease in fatty acid oxidation in liver. Fibrates also decrease thesynthesis and expression of triglycerides (Staels and Auwerx 1998).PPARα activators are also capable of correcting hyperglycemia andinsulin level. Fibrates also reduce adipose tissue mass through amechanism which is independent of food intake and leptin gene expression(Guerre-Millo, Gervois et al. 2000).

The therapeutic interest of PPARγ agonists has been widely investigatedin the treatment of type 2 diabetes (Spiegelman 1998). It has been shownthat PPARγ agonists restore insulin sensitivity in target tissues andreduce plasma glucose, lipid and insulin levels both in animal models oftype 2 diabetes and in humans (Ram V J 2003).

PPAR activation by ligands also plays a role in regulating theexpression of genes that participate in processes such as inflammation,angiogenesis, cell proliferation and differentiation, apoptosis and theactivities of iNOS, MMPase and TIMPs. Activation of PPARα inkeratinocytes results in a cessation of their proliferation andexpression of genes involved in differentiation (Komuves, Hanley et al.2000). The PPARs have anti-inflammatory properties because theynegatively interfere with transcription mechanisms involving othertranscription factors like NF-κB or transcriptional activators like(STAT) and AP-1 (Desvergne and Wahli 1999). Said anti-inflammatory andanti-proliferative properties make the PPARS (and particularly PPARα)interesting therapeutic targets for the treatment of diseases such asvascular occlusive diseases (atherosclerosis, etc.), hypertension,diseases related to neovascularization (diabetic retinopathy, etc.),inflammatory diseases (inflammatory bowel disease, psoriasis, etc.) andneoplastic diseases (carcinogenesis, etc.).

Free radicals play a role in a very large range of pathologies includingallergy, tumor initiation and promotion, cardiovascular diseases(atherosclerosis, ischemia), genetic and metabolic disorders (diabetes),infectious and degenerative diseases (Alzheimer's disease, Parkinson'sdisease, Prion, etc.) and ophthalmic disorders (Mates, Perez-Gomez etal. 1999).

Reactive oxygen species (ROS) are produced during normal cellfunctioning. ROS comprise the hydroxyl radicals (OH), superoxide anion(O₂ ⁻), hydrogen peroxide (H₂O₂) and nitric oxide (NO). Said species arevery labile and, due to their high chemical reactivity, constitute adanger to the biological functions of cells. They induce lipidperoxidation, oxidation of certain enzymes and very extensive oxidationof proteins leading to degradation thereof. Protection against lipidperoxidation is an essential process in aerobic organisms, becauseperoxidation products can cause DNA damage. Thus a deregulation ormodification of the equilibrium between the production, processing andelimination of radical species by natural antioxidant defenses leads tothe establishment of processes that are deleterious to the cell ororganism.

ROS are processed via an antioxidant system that comprises an enzymaticcomponent and a non-enzymatic component.

The enzymatic system is composed of several enzymes which have thefollowing characteristics:

-   -   Superoxide dismutase (SOD) destroys the superoxide radical by        converting it to peroxide. The peroxide in turn is acted upon by        another enzyme system. Low levels of SOD are continuously        produced by aerobic respiration. Three classes of SOD have been        identified in humans, each containing Cu, Zn, Fe, Mn, or Ni as        cofactor. The three forms of human SOD are distributed as        follows: a cytosolic Cu—Zn SOD, a mitochondrial Mn—SO and an        extracellular SOD.    -   Catalase is very efficient at converting hydrogen peroxide        (H₂O₂) to water and O₂. Hydrogen peroxide is enzymatically        catabolized in aerobic organisms. Catalase also catalyzes the        reduction of a variety of hydroperoxides (ROOH).    -   Glutathione peroxidase uses selenium as cofactor and catalyzes        the reduction of hydroperoxides (ROOH and H₂O₂) by using        glutathione, and thereby protects cells against oxidative        damage.

Non-enzymatic antioxidant defenses comprise molecules that aresynthesized or supplied in the diet.

Antioxidant molecules are present in different cell compartments.Detoxification enzymes for example eliminate free radicals and areessential to cell life. The three most important types of antioxidantcompounds are the carotenoids, vitamin C and vitamin E (Gilgun-Sherki,Melamed et al. 2001).

The inventors have shown that, in a surprising manner, the compounds ofthe invention have PPARα agonist activity and antioxidant properties.The inventive compounds are therefore capable of interfering with atleast two signalling pathways which are activated in particular ininflammation: cytokine production and free radical production. By actingsynergistically, the compounds of the invention constitute anadvantageous therapeutic means for treating pathologies related toinflammation (atherosclerosis, allergy, asthma, eczema, pruritus, etc.),neurodegeneration (Alzheimer's disease, Parkinson's disease, etc.),deregulations of lipid and/or glucose metabolism (diabetes,atherosclerosis, obesity, etc.), cell proliferation/differentiation(carcinogenesis, etc.) and disorders related to ageing (skin or centralnervous system, etc.).

The inventors have shown that the compounds according to the inventionconcurrently have PPAR activator, antioxidant and anti-inflammatoryproperties.

Thus, the invention concerns pharmaceutical compositions comprising atleast one substituted 1,3-diphenylprop-2-en-1-one derivative for thetreatment of pathologies related to inflammation, neurodegeneration,deregulations of lipid and/or glucose metabolism, cell proliferationand/or differentiation and/or skin or central nervous system ageing.

In particular, then, the invention has as its object a compositioncomprising, in a pharmaceutically acceptable support, at least onesubstituted 1,3-diphenylprop-2-en-1-one derivative represented byformula (I) below:

in which:

-   X1 represents a halogen or a —R1 group or a group corresponding to    the following formula: -G1-R1,-   X2 represents a hydrogen atom or a thionitroso group or a hydroxy    group or an unsubstituted alkyloxy group or an alkylcarbonyloxy    group or a thiol group or an alkylthio group or an alkylcarbonylthio    group, X2 can also represent an oxygen or sulfur atom bound to    carbon 3 of the propene chain, so as to form a derivative of the    type 2-phenyl-4H-1-benzopyran-4-one (this option is depicted in    formula (I) by dotted lines),-   X3 represents a —R3 group or a group corresponding to the following    formula: -G3-R3,-   X4 represents a halogen or a thionitroso group or a —R4 group or a    group corresponding to the following formula: -G4-R4,-   X5 represents a —R5 group or a group corresponding to the following    formula: -G5-R5,-   X6 is an oxygen atom or a nitrogen atom, in the case where X6 is a    nitrogen atom, it carries a hydrogen atom or a hydroxy group or an    alkyloxy group,-   R1, R3, R4, R5, which are the same or different, represent a    hydrogen atom or an alkyl group substituted or not by at least one    substituent which is part of group 1 or group 2 defined hereinbelow,-   G1, G3, G4, G5, which are the same or different, represent an oxygen    or sulfur atom,-   with at least one of the groups X1, X3, X4 or X5 corresponding to    the formula -G-R, and-   with at least one of the groups R1, R3, R4 or R5 present in the form    of an alkyl group containing at least one substituent from group 1    or group 2, said alkyl group being bound directly to the ring or    being associated with a group G according to the formula -GR,-   the substituents from group 1 are selected in the group consisting    of carboxy groups having the formula: —COOR₆ and carbamoyl groups    having the formula: —CONR₆R₇,-   the substituents from group 2 are selected in the group consisting    of sulfonic acid (—SO₃H) and sulfonamide groups having the formula:    —SO₂NR₆R₇-   with R₆ and R₇, which are the same or different, representing a    hydrogen atom or an alkyl group possibly substituted by at least one    group of type 1 or 2,-   the optical and geometrical isomers, racemates, tautomers, salts,    hydrates and mixtures thereof,-   with the exception of compounds represented by formula (I) in which:-   X₁, X₂, X₃ and X₅ each represent a hydrogen atom, X₆ represents an    oxygen atom and X₄ represents a group corresponding to the formula    —O—CR₈R₉—COOR₁₀, where R₈ and R₉, which are the same or different,    represent a C1 to C2 alkyl group (comprising one or two carbon    atoms), and R₁₀ represents a hydrogen atom or a C1 to C7 alkyl    group,-   X₂, X₃ and X₅ each represent a hydrogen atom, X₁ represents a    halogen atom or a R1 or -G1 R1 group, where R1 represents an    unsubstituted C1 to C2 alkyl group and G1 represents an oxygen atom,    X₆ represents an oxygen atom and X₄ represents a group corresponding    to the formula —O—CR₁₁R₁₂—COOR₁₀, where R₁₁ and R₁₂, which are the    same or different, represent a hydrogen atom or a C1 to C2 alkyl    group, and R₁₀ represents a hydrogen atom or a C1 to C7 alkyl group    (comprising one to seven carbon atoms), and-   X₂ represents a hydrogen atom and X, represents -G1R1 where G1    represents an oxygen atom and R1 represents CH2COOH.

Said composition can be used in particular for the treatment orprophylaxis of a pathology related to inflammation, neurodegeneration,deregulations of lipid and/or glucose metabolism, cell proliferationand/or differentiation and/or skin or central nervous system ageing.

The invention also encompasses a composition comprising the prodrugs ofcompounds represented by formula (I) which, after administration to asubject, are converted to compounds represented by formula (I) and/ormetabolites of compounds represented by formula (I) which displaysimilar therapeutic activity to compounds represented by formula (I),possibly in association with another therapeutic agent, for thetreatment or prophylaxis of a pathology related to inflammation,neurodegeneration, deregulations of lipid and/or glucose metabolism,cell proliferation and/or differentiation and/or skin or central nervoussystem ageing.

In the scope of the invention, the derivatives represented by formula(I) such as defined hereinabove can adopt a cis or trans conformation.An composition according to tha invention can thus comprise derivativescorresponding to the cis or trans conformation or a mixture thereof.

Advantageously, none of the groups X3, X4 and X5 represents a hydrogenatom. Compounds with formula (I) which meet this definition constitutecompounds of general formula (II).

Advantageously, one or two of the groups X3, X4 and X5 represent ahydrogen atom and XI is an unsubstituted alkyl group. Compounds withformula (I) which meet this definition constitute compounds of generalformula (III).

Advantageously, one or two of the groups X3, X4 and X5 represent ahydrogen atom and X2 is a thionitroso group or an alkylcarbonyloxy groupor a thiol group or an alkylthio group or an alkylcarbonylthio group, X2can also represent an oxygen or sulfur atom bound to carbon 3 of thepropene chain, so as to form a derivative of the type2-phenyl-4H-1-benzopyran-4-one (this option is depicted in formula (I)by dotted lines). Compounds with formula (I) which meet this definitionconstitute compounds of general formula (IV).

Advantageously, one or two of the groups X3, X4 and X5 represent ahydrogen atom and at least one of the groups X1, X2, X3, X4 or X5 is theGR form in which G is a sulfur atom. Compounds with formula (I) whichmeet this definition constitute compounds of general formula (V).

Advantageously, one or two of the groups X3, X4 and X5 represent ahydrogen atom and at least one of the groups X1, X3, X4 or X5 is the-G-R form in which G is an oxygen atom and R is an alkyl groupsubstituted by a substituent from group 1 where R6 is not a hydrogenatom. Compounds with formula (I) which meet this definition constitutecompounds of general formula (VI).

Advantageously, one or two of the groups X3, X4 and X5 represent ahydrogen atom and at least one of the groups X1, X3, X4 or X5 is the GRform in which G is an oxygen atom and R is an alkyl group substituted bya sulfonamide such as defined hereinabove. Compounds with formula (I)which meet this definition constitute compounds of general formula(VII).

Advantageously, X4 is a thionitroso group or a —R4 group or a groupcorresponding to the formula -G4-R4. Derivatives having formula (I) inwhich X4 meets this definition constitute derivatives represented bygeneral formula (VIII) in which G4 and R4 are such as definedhereinabove.

Advantageously, X2 is a thionitroso group or a hydroxy group or analkyloxy group or a thiol group or an alkylthio group. Derivativeshaving formula (I) in which X2 meets this definition constitutederivatives represented by general formula (IX).

Other advantageous derivatives represented by formula (I) of theinvention have a general formula (X) such that X4 is a thionitroso groupor a —R4 group or a group corresponding to the formula -G4-R4 and X2 isa thionitroso group or a hydroxy group or an alkyloxy group or a thiolgroup or an alkylthio group, G4 and R4 being such as definedhereinabove.

Other advantageous derivatives represented by formula (I) of theinvention have a general formula (XI) such that X1 represents a —R1group or a group corresponding to the formula -G1-R1, with R1 being analkyl group substituted by a substituent which is part of group 1 and G1and the substituent from group 1 being such as defined hereinabove.

More preferably, another object of the invention concerns derivativesrepresented by formula (XI) such as described hereinabove, characterizedin that X1 is a -G1-R1 group.

Even more preferably, another object of the invention concernsderivatives represented by formula (XI) such as described hereinabove,characterized in that X1 is a -G1-R1 group in which G1 is an oxygenatom.

Other advantageous derivatives represented by formula (I) of theinvention have a general formula (XII) such that X1 represents a —R1group or a group corresponding to the formula -G1-R1, with R1 being analkyl group substituted by a substituent which is part of group 2 and G1and the substituent from group 2 being such as defined hereinabove.

Other advantageous derivatives represented by formula (I) of theinvention have a general formula (XIII) such that X3 represents a —R3group or a group corresponding to the formula -G3-R3, with R3 being analkyl group substituted by a substituent which is part of group 1 and G3and the substituent from group 1 being such as defined hereinabove.

Other advantageous derivatives represented by formula (I) of theinvention have a general formula (XIV) such that X3 represents a —R3group or a group corresponding to the formula -G3-R3, with R3 being analkyl group substituted by a substituent which is part of group 2 and G3and the substituent from group 2 being such as defined hereinabove.

Other advantageous derivatives represented by formula (I) of theinvention have a general formula (XV) such that X4 represents a —R4group or a group corresponding to the formula -G4-R4, with R4 being analkyl group substituted by a substituent which is part of group 1 and G4and the substituent from group 1 being such as defined hereinabove.

More preferably, another object of the invention concerns derivativesrepresented by formula (XV) such as described hereinabove, characterizedin that X4 is a -G4-R4 group.

Even more preferably, another object of the invention concernsderivatives represented by formula (XV) such as described hereinabove,characterized in that X4 is a -G4-R4 group in which G4 is an oxygenatom.

Even more preferably, another object of the invention concernsderivatives represented by formula (XV) such as described hereinabove,characterized in that X4 is a -G4-R4 group in which G4 is an oxygenatom, and X3 or X5 respectively represent R3 or G3R3, on the one hand,and R5 or G5R5, on the other hand, with R3 or R5 being alkyl groupscarrying a substitutent from group 1, said substituent from group 1being such as defined hereinabove.

Other advantageous derivatives represented by formula (I) of theinvention have a general formula (XVI) such that X4 represents a —R4group or a group corresponding to the formula -G4-R4 with R4 being analkyl group substituted by a substituent which is part of group 2 and G4the substituent from group 2 being such as defined hereinabove.

Other advantageous derivatives represented by formula (I) of theinvention have a general formula (XVII) such that X1 represents ahalogen.

Other advantageous derivatives represented by formula (I) of theinvention have a general formula (XVIII) such that X1 represents a —R1group with R1 being a C1 to C4 alkyl group substituted or not by atleast one substituent which is part of group 1 or group 2 definedhereinabove.

Other advantageous derivatives represented by formula (I) of theinvention have a general formula (XIX) such that X1 represents a -G1R1group with R1 being a C1 to C3 alkyl group substituted or not by atleast one substituent which is part of group 1 or group 2 definedhereinabove.

Other advantageous derivatives represented by formula (I) of theinvention have a general formula (XX) such that X1 represents a —R1group with R1 being a C5 to C24 alkyl group substituted or not by atleast one substituent which is part of group 1 or group 2 definedhereinabove.

Other advantageous derivatives represented by formula (I) of theinvention have a general formula (XXI) such that X1 represents a -G1R1group with R1 being a C4 to C24 alkyl group substituted or not by atleast one substituent which is part of group 1 or group 2 definedhereinabove.

Another object of the invention concerns derivatives represented byformula (I) in which X1, X3, X4 or X5 represent OC(CH3)2COOR6 with R6being such as defined hereinabove.

Another object of the invention concerns derivatives represented byformula (I) in which X1, X3, X4 or X5 represent SC(CH3)2COOR6 with R6being such as defined hereinabove.

According to the invention, the term “alkyl” designates a saturatedhydrocarbon function, linear, branched or cyclic, halogenated or not,having more particularly from 1 to 24, preferably 1 to 10, carbon atomssuch as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl,pentyl, neopentyl, n-hexyl. Groups containing one or two carbon atoms orcontaining from two to seven carbon atoms are particularly preferred.Methyl and ethyl groups are more particularly preferred.

The term thionitroso refers to a nitroso group bound to the aromaticring through a sulfur atom.

The term halogen represents a chlorine atom or a bromine atom or aniodine atom or a fluorine atom.

The term alkyloxy designates an alkyl chain bound to the ring by anoxygen atom. The alkyl chain is defined earlier.

The term alkylthio refers to an alkyl chain bound to the aromatic ringby a sulfur atom (thioether bond). The alkyl chain is defined earlier.

According to a particular embodiment of the invention, preferredcompounds are indicated below with their corresponding formulas:

-   1-[2-hydroxy-4-chlorophenyl]-3-[4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one    and 1-[2-hydroxy-4-chlorophenyl]-3-[4-isopropyloxycarbonyl    dimethylmethyloxyphenyl]prop-2-en-1-one

-   1-[2-hydroxyphenyl]-3-[4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one    and    1-[2-hydroxyphenyl]-3-[4-isopropyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one:

-   1-[2-methylcarbonyloxyphenyl]-3-[4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one    and 1-[2-methylcarbonyloxyphenyl]-3-[4-isopropyloxycarbonyldimethyl    methyloxyphenyl]prop-2-en-1-one:

-   1-[2-hydroxyphenyl]-3-[4-carboxydimethylmethyloxyphenyl]-1-hydroxyiminoprop-2-ene    and    1-[2-hydroxyphenyl]-3-[4-isopropyloxycarbonyldimethylmethyloxyphenyl]-1-hydroxyiminoprop-2-ene:

-   1-[2-hydroxy-4-carboxydimethylmethyloxyphenyl]-3-[3,    5-ditertbutyl-4-hydroxyphenyl]prop-2-en-1-one,    1-[2-hydroxy-4-ethyloxycarbonyl    dimethylmethyloxyphenyl]-3-[3,5-ditertbutyl-4-hydroxyphenyl]prop-2-en-1-one    and    1-[2-hydroxy-4-ethoxycarbonyldimethylmethyloxyphenyl]-3-[3,5-ditertbutyl-4-hydroxyphenyl]prop-2-en-1-one:

-   1-[2-hydroxyphenyl]-3-[3-carboxydimethylmethyloxy-4-hydroxy-5-tertbutylphenyl]prop-2-en-1-one    and    1-[2-hydroxyphenyl]-3-[3-isopropyloxycarbonyldimethylmethyloxy-4-hydroxy-5-tertbutylphenyl]pro    p-2-en-1-one:

-   1-[2-hydroxy-4-chlorophenyl]-3-[3-carboxydimethylmethyloxy-4-hydroxy-5-tertbutylphenyl]prop-2-en-1-one    and    1-[2-hydroxy-4-chlorophenyl]-3-[3-isopropyloxycarbonyldimethylmethyloxy-4-hydroxy-5-tertbutylphenyl]prop-2-en-1-one:

-   1-[2-hydroxyphenyl]-3-[3-carboxydimethylmethyl-4-hydroxy-5-tertbutylphenyl]prop-2-en-1-one    and    1-[2-hydroxyphenyl]-3-[3-isopropyloxycarbonyldimethylmethyl-4-hydroxy-5-tertbutylphenyl]prop-2-en-1-one    (:

-   1-[2-hydroxy-4-chlorophenyl]-3-[3-carboxydimethylmethyl-4-hydroxy-5-tertbutylphenyl]prop-2-en-1-one    and    1-[2-hydroxy-4-chlorophenyl]-3-[3-isopropyloxycarbonyldimethylmethyl-4-hydroxy-5-tertbutylphenyl]prop-2-en-1-one:

-   1-[2-hydroxy-4-chlorophenyl]-3-[3,5-dimethoxy-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one    and    1-[2-hydroxy-4-chlorophenyl]-3-[3,5-dimethoxy-4-isopropyloxycarbonyldimethylmethytoxy    phenyl]prop-2-en-1-one:

-   1-[2-hydroxyphenyl]-3-[3,5-dimethoxy-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one    and    1-[2-hydroxyphenyl]-3-[3,5-dimethoxy-4-isopropyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one:

-   1-[2-hydroxy-4-carboxydimethylmethyloxyphenyl]-3-[3,5-dimethoxy-4-hydroxyphenyl]prop-2-en-1-one    and    1-[2-hydroxy-4-isopropyloxycarbonyldimethylmethyloxyphenyl]-3-[3,5-di-methoxy-4-hydroxyphenyl]prop-2-en-1-one

-   1-[2-hydroxy-4-chlorophenyl]-3-[3,4-dihydroxy-5-carboxydimethylmethyloxyphenyl]prop-2-en-1-one    and    1-[2-hydroxy-4-chlorophenyl]-3-[3,4-dihydroxy-5-isopropyloxycarbonyldimethylmethyloxyphenyl]-2-propen-1-one:

-   1-[2-hydroxy-4-carboxydimethylmethyloxyphenyl]-3-[3,5-dimethyl-4-hydroxyphenyl]prop-2-en-1-one    and 1-[2-hydroxy-4-isopropyloxycarbonyl    dimethylmethyloxyphenyl]-3-[3,5-dimethyl-4-hydroxyphenyl]prop-2-en-1-one:

-   1-[2-hydroxy-4-chlorophenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyl    oxyphenyl]prop-2-en-1-one and    1-[2-hydroxy-4-chlorophenyl]-3-[3,5-dimethyl-4-isopropyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one:

-   and    1-[2-hydroxyphenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one    and    1-[2-hydroxyphenyl]-3-[3,5-dimethyl-4-isopropyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one:

-   and    1-[2-hydroxyphenyl]-3-[3-carboxydimethylmethyloxyphenyl]prop-2-en-1-one    and    1-[2-hydroxyphenyl]-3-[3-isopropyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one:

-   1-[2-hydroxyphenyl]-3-[4-carboxydimethylmethylthiophenyl]prop-2-en-1-one    and    1-[2-hydroxyphenyl]-3-[4-isopropyloxycarbonyldimethylmethylthiophenyl]prop-2-en-1-one:

-   1-[2-mercapto-4-methyloxyphenyl]-3-[4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one    and    1-[2-mercapto-4-methyloxyphenyl]-3-[4-isopropyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one:

-   1-[4-heptylphenyl]-3-[3-methyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one    and    1-[4-heptylphenyl]-3-[3-methyl-4-isopropyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one:

-   1-[2-hydroxy-4-carboxydimethylmethyloxyphenyl]-3-[3,5-dibromo-4-hydroxyphenyl]prop-2-en-1-one,-   1-[2-hydroxy-4-carboxydimethylmethyloxyphenyl]-3-[3-hydroxyphenyl]prop-2-en-1-one,-   1-[2-hydroxy-4-carboxydimethylmethyloxyphenyl]-3-[4-methylthiophenyl]prop-2-en-1-one,-   1-[2-hydroxy-4-carboxydimethylmethyloxyphenyl]-3-[4-methylthiophenyl]prop-2-en-1-one,-   1-[2,4-dihydroxyphenyl]-3-[4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one,-   1-[2-hydroxyphenyl]-3-[4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one,-   1-[4-chlorophenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one,-   1-[4-chlorophenyl]-3-[3,5-dimethyl-4-isopropyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one,-   1-[4-chlorophenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one,-   1-[2-hydroxy-4-carboxydimethylmethyloxyphenyl]-3-[4-chlorophenyl]prop-2-en-1-one,-   1-[2-hydroxyphenyl]-3-[4-carboxydimethylmethylthiophenyl]prop-2-en-1-one,-   1-[4-chloro-2-hydroxyphenyl]-3-[4-carboxydimethylmethylthiophenyl]prop-2-en-1-one,-   1-[4-carboxydimethylmethyloxyphenyl]-3-[3,5-dimethyl4-hydroxyphenyl]prop-2-en-1-one,-   1-[4-methylthiophenyl]-3-[4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one,-   1-[4-carboxydimethylmethyloxyphenyl]-3-[4-chlorophenyl]prop-2-en-1-one,-   1-[4-carboxydimethylmethylthiophenyl]-3-[4-methylthiophenyl]prop-2-en-1-one,-   1-[2-hydroxy-4-bromophenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one,-   1-[4-carboxydimethylmethyloxyphenyl]-3-[4-methylthiophenyl]prop-2-en-1-one,-   1-[4-methylthiophenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one,-   1-[4-methylthiophenyl]-3-[3,5-dimethyl-4-isopropyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one,-   1-[4-methylthiophenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one,-   1-[2-methoxyphenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one,-   1-[2-methoxyphenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one,-   1-[4-hexyloxyphenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one,-   1-[4-hexyloxyphenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one,-   2-(3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl)-7-chloro-4H-1-benzopyran-4-one,-   2-(3,5-dimethyl-4-carboxydimethylmethyloxyphenyl)-7-chloro-4H-1-benzopyran-4-one,-   1-[2-methyloxy-4-chlorophenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one,-   1-[2-methyloxy-4-chlorophenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one,-   1-[4-heptylphenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one,-   1-[4-heptylphenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one,-   1-[4-bromophenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one,-   1-[4-bromophenyl]-3-[3,5-dimethyl-4-carboxy    dimethylmethyloxyphenyl]prop-2-en-1-one,-   1-[2-hydroxyphenyl]-3-[3,5-dimethyl-4-isopropyloxycarbonyl    dimethylmethyloxyphenyl]prop-2-en-1-one

The invention thus concerns the use of at least one compound representedby formula (I) such as defined hereinabove and in particular one of theadvantageous or preferred compounds for preparing a pharmaceuticalcomposition for the preventive or preferably curative treatment of apathology related to inflammation, neurodegeneration, deregulations oflipid and/or glucose metabolism, cell proliferation and/ordifferentiation and/or skin or central nervous system ageing, such asallergy, asthma, eczema, psoriasis, pruritus, Alzheimer's disease,Parkinson's disease, diabetes, atherosclerosis, obesity, carcinogenesis,etc.

The invention also provides a method for preparing compounds orderivatives represented by formula (I).

The method of the invention comprises contacting in a basic medium or inan acidic medium at least one compound represented by formula (A) withat least one compound represented by formula (B), formulas (A) and (B)being:

formulas in which X1, X2, X3, X4 and X5 are such as defined hereinabove.

The conditions for carrying out said reaction in acidic or basic mediumare within reach of those skilled in the art and can vary widely.

Said two compounds are advantageously contacted in stoichiometricproportions. Contact is preferably carried out at room temperature(between approximately 18° C. and 25° C.) and at atmospheric pressure.

In basic medium, the reaction is preferably carried out in the presenceof a strong base, such as an alkaline earth metal hydroxide, like sodiumhydroxide or an alkaline metal alcoholate like sodium ethylate.

In acidic medium, the reaction is preferably carried out in the presenceof a strong acid, such as hydrochloric acid.

The reaction scheme may be depicted as follows:

The synthesis in basic medium may be carried out in the followingmanner:

-   One molar equivalent of ketone (compound (A)) and one molar    equivalent of aldehyde (compound (B)) are solubilized in a    hydroalcoholic solution of 20 molar equivalents of sodium hydroxide.    The mixture is stirred for approximately 18 hours at room    temperature (between 18° C. and 25° C.). The medium is then    acidified (in particular to a pH of approximately 2) in particular    with hydrochloric acid-   The expected substituted 1,3-diphenylprop-2-en-1-one can be obtained    by precipitation or solid/liquid extraction after evaporation of the    reaction medium. It can then be purified by silica gel    chromatography or by crystallization.-   The synthesis in acidic medium may be carried out in the following    manner:-   One molar equivalent of ketone (compound (A)) and one molar    equivalent of aldehyde (compound (B)) are solubilized in an ethanol    solution saturated with gaseous hydrochloric acid. The mixture is    stirred at room temperature for approximately 6 hours, the solvent    is eliminated, in particular by vacuum evaporation. The substituted    1,3-diphenylprop-2-en-1-one is purified, in particular by    chromatography on silica gel.

The invention thus concerns the use of a compound or derivative such asdefined hereinabove for preparing a pharmaceutical composition forpracticing a method of treatment or prophylaxis of the human or animalbody.

The pharmaceutical compositions or compounds represented by formula (I)according to the invention are advantageously used for the treatment orprophylaxis of pathologies related to inflammation, neurodegeneration,deregulations of lipid and/or glucose metabolism, cell proliferationand/or differentiation and/or skin or central nervous system ageing andmore particularly of one or more allergies, asthma, eczema, psoriasis,pruritus, Alzheimer's disease, Parkinson's disease, diabetes,atherosclerosis, obesity, carcinogenesis, etc. In fact, it was foundsurprisingly that compounds represented by formula (I) have advantageouspharmacological properties as antioxidants and activators of PPARα andPPARγ.

In the case where the composition of the invention is intended for thetreatment or prophylaxis of a pathology related to neurodegeneration,deregulations of lipid and/or glucose metabolism, cell proliferationand/or differentiation and/or skin or central nervous system ageing, thecompounds represented by formula (I) can optionally include those havingformula (I) in which X₂ represents a hydrogen atom and X₁ represents-G1R1 where G1 represents an oxygen atom and R1 represents CH2COOH.Preferably, however, said compounds are excluded.

In the case where the composition of the invention is intended for thetreatment or prophylaxis of a pathology related to inflammation,neurodegeneration, cell proliferation and/or differentiation and/or skinor central nervous system ageing and more particularly of one or moreallergies, asthma, eczema, psoriasis, pruritus, Alzheimer's disease,Parkinson's disease or carcinogenesis, the compounds represented byformula (I) can optionally include those having formula (I) in which:

-   X₁, X₂, X₃ and X₅ each represent a hydrogen atom, X₆ represents an    oxygen atom and X₄ represents a group corresponding to the formula    —O—CR₈R₉—COOR₁₀, where R₈ and R₉, which are the same or different,    represent a C1 to C2 alkyl group (comprising one or two carbon    atoms), and R₁₀ represents a hydrogen atom or a C1 to C7 alkyl    group,-   X₂, X₃ and X₅ each represent a hydrogen atom, X₁ represents a    halogen atom or a R1 or -G1R1 group, where R1 represents an    unsubstituted C1 to C2 alkyl group and G1 represents an oxygen atom,    X₆ represents an oxygen atom and X₄ represents a group corresponding    to the formula —O—CR₁₁R₁₂—COOR₁₀, where R₁₁ and R₁₂, which are the    same or different, represent a hydrogen atom or a C1 to C2 alkyl    group, and R₁₀ represents a hydrogen atom or a C1 to C7 alkyl group    (comprising one to seven carbon atoms).

The invention also concerns a method for treating pathologies related toinflammation, neurodegeneration, deregulations of lipid and/or glucosemetabolism, cell proliferation and/or differentiation and/or skin orcentral nervous system ageing and more particularly of one or moreallergies, asthma, eczema, psoriasis, pruritus, Alzheimer's disease,Parkinson's disease, diabetes, atherosclerosis, obesity, carcinogenesis,comprising administering to a subject, particularly human, an effectivedose of a compound represented by general formula (I) or of apharmaceutical composition such as defined hereinabove.

The pharmaceutical compositions according to the inventionadvantageously comprise one or more pharmaceutically acceptableexcipients or vehicles. Examples include saline, physiological,isotonic, buffered solutions and the like, compatible withpharmaceutical use and known to those skilled in the art. Thecompositions may contain one or more agents or vehicles selected in thegroup consisting of dispersants, solubilizers, stabilizers,preservatives, and the like. Agents or vehicles that can be used in theformulations (liquid and/or injectable and/or solid) are in particularmethylcellulose, hydroxymethylcellulose, carboxymethylcellulose,polysorbate 80, mannitol, gelatin, lactose, plant oils, acacia, and thelike. The compositions may be formulated as suspensions for injection,gels, oils, tablets, suppositories, powders, capsules, gelules, and thelike, possibly by means of pharmaceutical forms or devices ensuringprolonged and/or delayed release. For this type of formulation, an agentsuch as cellulose, carbonates or starch is advantageously used.

The compounds or compositions of the invention may be administered indifferent ways and in different forms. For instance, they may beadministered by the oral or systemic route, such as for example by theintravenous, intramuscular, subcutaneous, transdermal, intra-arterialroute, etc. For injections, the compounds are generally formulated asliquid suspensions, which can be injected through syringes or byinfusion, for example. It is understood that the injection rate and/orthe injected dose may be adapted by those skilled in the art accordingto the patient, the pathology, the method of administration, etc.Typically, the compounds are administered at doses ranging from 1 μg to2 g per administration, preferably from 0.1 mg to 1 g peradministration. The administrations may be given daily or repeatedseveral times a day, as the case may be. Moreover, the inventivecompositions may additionally comprise other active ingredients oragents.

Other aspects and advantages of the invention will become apparent inthe following examples, which are given for purposes of illustration andnot by way of limitation.

LEGENDS OF FIGURES

FIG. 1-1, 1-2, 1-3: Evaluation of the antioxidant properties of compound2, compound 3, compound 12, compound 14 and compound 17 on LDL oxidationby copper (Cu).

FIG. 1-1 shows the results of the experiment measuring formation ofconjugated dienes over time. It can be seen that incubation of LDL withthe test compounds at a concentration of 10⁻⁴M delayed conjugated dieneformation. The lag phase was 111 minutes for copper alone as comparedwith a lag phase of 132, 145, 134 and 203 minutes, respectively, whenLDL were incubated with compound 3, compound 12, compound 14, compound17. The lag phase was more than 480 minutes when LDL were incubated withcompound 2. This lag in the formation of conjugated dienes ischaracteristic of antioxidants.

FIG. 1-2 shows the rate of diene formation after different treatments.Incubation of the compounds with LDL in the presence of copper slowedthe rate of conjugated diene formation. This rate was 2 nmol/min/mg ofLDL with copper alone, 1.7 nmol/min/mg of LDL when LDL were incubated inthe presence of 10⁻⁴M compound 17, and not determined for compound 2 at10⁻⁴M (not measurable because too low).

FIG. 1-3 represents the maximum amount of conjugated dienes formed overtime. Incubation of LDL with copper led to formation of 348 nmol ofconjugated dienes per mg of LDL; incubation with compound 2 at 10⁻⁴M ledto an 84% decrease in conjugated diene formation (54.4 nmol per mg ofLDL). In the presence of compounds 3 and 17, conjugated diene formationwas respectively 303 and 327 nmol per mg of LDL.

FIG. 1-4, 1-5, 1-6: Evaluation of the antioxidant properties of compound18, compound 19, compound 21 and compound 22 on LDL oxidation by copper(Cu).

FIG. 1-4 shows that incubation of LDL with the test compounds at aconcentration of 10⁻⁴M delayed conjugated diene formation. The lag phasewas 178 minutes for copper alone as compared with a lag phase of 241,182 and 241 minutes (from the experimental determination), respectively,when LDL were incubated with compound 18, compound 19, or compound 22.The lag phase was more than 480 minutes when LDL were incubated withcompound 21. This lag in the formation of conjugated dienes ischaracteristic of antioxidants.

FIG. 1-5 shows the rate of diene formation after different treatments.The rate of formation of conjugated dienes was 1.6 nmol/min/mg of LDLwith copper alone, 1.4 nmol/min/mg of LDL when LDL were incubated in thepresence of compound 18 at 10⁻⁴M, 1.3 nmol/min/mg of LDL when LDL wereincubated in the presence of compounds 22, and not determined forcompound 21 at 10⁻⁴M (not measurable because too low).

FIG. 1-6 represents the maximum amount of conjugated dienes formed overtime. Incubation of LDL with copper led to formation of 353 nmol ofconjugated dienes per mg of LDL. Incubation with compound 21 at 10⁻⁴Minhibited conjugated diene formation. Conjugated diene formation wasrespectively 305, 345 and 345 nmol per mg of LDL in the presence ofcompounds 18, 19 and 22.

FIG. 1-7. 1-8: Evaluation of the antioxidant properties of compound 25and compound 28 on LDL oxidation by copper (Cu).

FIG. 1-7 shows the results of the experiment measuring formation ofconjugated dienes over time. It can be seen that incubation of LDL withthe test compounds at a concentration of 10⁻⁴M delayed conjugated dieneformation. The lag phase was 82 minutes for copper alone as comparedwith a lag phase of 120 and 135 minutes (from the experimentaldetermination), respectively, when LDL were incubated with compound 25and compound 29.

FIG. 1-8 represents the maximum amount of conjugated dienes formed overtime. Incubation of LDL with copper led to formation of 393 nmol ofconjugated dienes per mg of LDL. In the presence of compound 25 thisvalue was 378 nmol per mg of LDL.

FIGS. 1-9, 1-10, 1-11: Evaluation of the antioxidant properties ofcompound 31, compound 33 and compound 35 on LDL oxidation by copper(Cu).

FIG. 1-9 shows the results of the experiment measuring formation ofconjugated dienes over time. It can be seen that incubation of LDL withthe test compounds at a concentration of 10⁻⁴M delayed conjugated dieneformation. The lag phase was 80 minutes for copper alone as comparedwith a lag phase of 139, 247 and 149 minutes (from the experimentaldetermination), respectively, when LDL were incubated with compound 31,compound 33, and compound 35. This lag in the formation of conjugateddienes is characteristic of antioxidants.

FIG. 1-10 shows the rate of diene formation after different treatments.Incubation of the compounds with LDL in the presence of copper slowedthe rate of conjugated diene formation. This rate was 1.9 nmol/min/mg ofLDL with copper alone, 1.6 nmol/min/mg of LDL when LDL were incubated inthe presence of compound 31 at 10⁻⁴M, 0.8 nmol/min/mg of LDL when LDLwere incubated in the presence of compound 33 and 1.5 nmol/min/mg of LDLwhen LDL were incubated in the presence of compound 35.

FIG. 1-11 represents the maximum amount of conjugated dienes formed overtime. Incubation of LDL with copper led to formation of 298 nmol ofconjugated dienes per mg of LDL, as compared with 257 nmol per mg of LDLin the presence of compound 33.

FIG. 1-12, 1-13, 1-14: Evaluation of the antioxidant properties ofcompound 37, compound 38 and compound 41 on LDL oxidation by copper(Cu).

FIG. 1-12 shows the results of the experiment measuring formation ofconjugated dienes over time. It can be seen that incubation of LDL withthe test compounds at a concentration of 10⁻⁴M delayed conjugated dieneformation. The lag phase was 120 minutes for copper alone as comparedwith a lag phase of 196, 284 and 411 minutes (from the experimentaldetermination), respectively, when LDL were incubated with compound 37,compound 38, and compound 41.

FIG. 1-13 shows the rate of diene formation after different treatments.Incubation of the compounds with LDL in the presence of copper slowedthe rate of conjugated diene formation. This rate was 1.8 nmol/min/mg ofLDL with copper alone, 1.49 nmol/min/mg of LDL when LDL were incubatedin the presence of compounds 37 at 10⁻⁴M, 0.71 nmol/min/mg of LDL whenLDL were incubated in the presence of compounds 38 and 0.54 nmol/min/mgof LDL when LDL were incubated in the presence of compounds 41.

FIG. 1-14 represents the maximum amount of conjugated dienes formed overtime. Incubation of LDL with copper led to formation of 372 nmol ofconjugated dienes per mg of LDL, as compared with 338 nmol per mg ofLDL, 244 nmol per mg of LDL, and 71 nmol per mg of LDL in the presenceof compounds 37, 38 and 41, respectively.

The lag phase in the formation of conjugated dienes, the reduction inthe rate of diene formation and the decrease in the total amount ofdienes formed are characteristics of antioxidants.

FIGS. 2-1, 2-2, 2-3, 2-4, 2-5, 2-6: Evaluation of PPARα agonistproperties of the inventive compounds in the PPARα/Gal4 transactivationsystem.

RK13 cells were incubated with the different compounds at concentrationsof 10, 30 and 100 μM or 1, 10 and 100 μM for 24 hours. The results areexpressed as the induction factor (luminescent signal relative tountreated cells) after the different treatments. The higher theinduction factor the more potent the PPARα agonist activity.

FIG. 2-1: The results show the induction factors for compound 3,compound 4, compound 7, compound 8 and compound 9. The values of theseinduction factors are given in Table 2-1.

TABLE 2-1 Induction Compound Concentration factor Cp3 10 μM 30.12 30 μM27.27 100 μM 25.84 Cp4 10 μM 3.99 30 μM 22.15 100 μM 61.07 Cp7 10 μM36.48 30 μM 50.37 100 μM 37.84 Cp8 10 μM 0.62 30 μM 1.27 100 μM 9.98 Cp910 μM 2.11 30 μM 5.00 100 μM 28.19

The results show that compound 3 produced a maximum 27-fold induction ata concentration of 30 μM, compound 4 had a maximum induction factor of60 at 100 μM, 22 at 30 μM and 4 at 10 μM. Compound 7 had a maximuminduction factor of 50 at 100 μM. Compound 8 activated the system with amaximum induction factor of 10 at 100 μM. Compound 9 had an inductionfactor of 28 at 100 μM, the highest concentration

FIG. 2-2: The results show the induction factors for compound 11,compound 12, compound 13, compound 14 and compound 17. The values ofthese induction factors are given in Table 2-2.

TABLE 2-2 Induction Compound Concentration factor Cp11 1 μm 1.20 10 μm1.39 100 μm 10.19 Cp12 1 μm 1.12 10 μm 8.45 100 μm 22.54 Cp13 1 μm 1.2010 μm 1.10 100 μm 1.5 Cp14 1 μm 1.25 10 μm 1.36 100 μm 1.38 Cp17 1 μm79.76 10 μm 85.69 100 μm 13.80

The results show that compound 11 produced a maximum 10-fold inductionat a concentration of 100 μM, compound 12 had a maximum induction factorof 22 at 100 μM, 8 at 30 μM and 1 at 10 μM. Compounds 13 and 14 hadinduction factors comprised between 1.1 and 1.5 at the differentconcentrations tested. Compound 17 activated the system with a maximuminduction factor of 85 at 10 μM and a minimum induction factor of 13.8at the 100 μM concentration.

FIG. 2-3 The results show the induction factors for compound 19,compound 20, compound 21 and compound 22. The values of these inductionfactors are given in Table 2-3.

TABLE 2-3 Induction Compound Concentration factor Cp19 1 μm 1.20 10 μm15.62 100 μm 0.07 Cp20 1 μm 21.50 10 μm 53.45 100 μm 1.22 Cp21 1 μm 0.7810 μm 1.10 100 μm 22.80 Cp22 1 μm 2.40 10 μm 49.49 100 μm 2.73

The results show that compound 19 produced a maximum 15.6-fold inductionat 10 μM, compound 20 had a maximum induction factor of 53 at 10 μM.Compound 21 had induction factors comprised between 0.8 and 22 at thedifferent concentrations tested. Compound 22 activated the system with amaximum induction factor of 50 at the 10 μM concentration.

FIG. 2-4 The results show the induction factors for compounds 23, 24,25, 26 and 29. The values of these induction factors are given in Table2-4.

TABLE 2-4 Induction Compound Concentration factor Cp23 1 μm 1.55 10 μm3.67 100 μm 0.12 Cp24 1 μm 2.06 10 μm 11.62 100 μm 0.00 Cp25 1 μm 13.4810 μm 21.03 100 μm 7.01 Cp26 1 μm 1.75 10 μm 7.85 100 μm 1.08 Cp29 1 μm28.36 10 μm 25.26 100 μm 0.27

Compound 23 had a maximum induction factor of 3.6 at 10 μM, compound 24had a maximum induction factor of 11 at 10 μM. Compound 25 activated thesystem with induction factors comprised between 7 and 21 according tothe concentrations tested. Compound 26 had a maximum induction factor of7.8 for the 10 μM concentration, compound 29 had induction factors of 28and 25 at 1 and 10 μM, respectively.

FIG. 2-5 The results show the induction factors for compound 31 andcompound 33. The values of these induction factors are given in Table2-5.

TABLE 2-5 Induction Compound Concentration factor Cp31 1 μm 3.77 10 μm15.52 100 μm 1.21 Cp33 1 μm 22.05 10 μm 44.52 100 μm 77.62

Compound 31 activated the system with an induction factor of 15.5 at theconcentration of 10 μM. The induction factors for compound 33 were 22,44 and 77 for the 1, 10 and 100 μM concentrations, respectively.

FIG. 2-6: The results show the induction factors for compounds 37, 38and 41. The values of these induction factors are given in Table 2-6.

TABLE 2-6 Induction Compound Concentration factor Cp37 1 μm 24.55 10 μm27.83 100 μm 0.02 Cp38 1 μm 14.70 10 μm 22.22 100 μm 0.311 Cp41 1 μm34.61 10 μm 31.18 100 μm 3.39

The maximum induction factors for compounds 37, 38 and 41 were 27, 22and 31, respectively, at the 10 μm concentration.

These results demonstrate that the inventive compounds tested exhibitPPARγ ligand activity and thus enable the transcriptional activationthereof.

FIG. 2-7: Evaluation of PPARγ agonist properties of the inventivecompounds in the PPARγ/Gal4 transactivation system.

RK13 cells were incubated with the different compounds at concentrationsof 1, 10 and 100 μM for 24 hours. The results are expressed as theinduction factor (luminescent signal relative to untreated cells) afterthe different treatments. The higher the induction factor the morepotent the PPARγ agonist activity.

The results in the figure show the induction factors for compound 17,compound 33, and compound 29. The values of these induction factors aregiven in Table 2-7.

TABLE 2-7 Induction Compound factor Cp17 1 μm 15.37 10 μm 24.92 100 μm6.13 Cp33 1 μm 15.65 10 μm 33.90 100 μm 45.58 Cp29 1 μm 17.05 10 μm33.89 100 μm 0.01

The results show that compound 17 had a maximum induction factor of 25at 10 μM. Compound 33 had a maximum induction factor of 45.6 at 100 μMand compound 29 of 33.9 at 10 μM.

These results demonstrate that the inventive compounds tested exhibitPPARγ ligand activity and thus enable the transcriptional activationthereof.

FIGS. 3-1. 3-2, 3-3, 3-4: Evaluation of the effect of compound 7 andcompound 17 on metabolism of triglycerides and cholesterol.

FIGS. 3-1, 3-2, 3-3 and 3-4 illustrate the effects of treatment withcompounds 7 and 17 on triglyceride and cholesterol metabolism in ApoE2/E2 transgenic mice. Animals were treated by gavage with each compoundat a dose of 200 mg/kg for 7 days.

FIGS. 3-1 and 3-2 illustrate the decrease in plasma concentrations oftriglycerides and cholesterol induced by compounds 7 and 17.

FIGS. 3-3 and 3-4 show triglyceride and cholesterol distribution inlipoparticles evaluated by exclusion chromatography. A typicaldistribution of triglycerides and cholesterol is observed principally inlarge sized lipoparticles. It can also be seen that treatment withcompounds 7 and 17 decreased the triglycerides and cholesterol in thislipoparticle subfraction.

FIGS. 3-5, 3-6. 3-7. 3-8:

FIGS. 3-5, 3-6, 3-7 and 3-8 illustrate the effects of treatment withcompound 29 according to the invention on triglyceride and cholesterolmetabolism in Apo E2/E2 transgenic mice. Animals were treated withcompound 29 at the following doses: 200, 50, 12.5 and 3.15 mg/kg/day for8 days.

FIGS. 3-5 and 3-6 illustrate the dose-dependent decrease in plasmatriglyceride and cholesterol levels with a greater decrease withincreasing doses of compound 29.

FIGS. 3-7 and 3-8 show triglyceride and cholesterol distribution inlipoparticles evaluated by exclusion chromatography. A typicaldistribution of triglycerides and cholesterol is observed principally inlarge sized lipoparticles. A decrease in triglycerides and cholesterolin this lipoparticle subfraction can also be seen.

FIGS. 3-9. 3-10. 3-11 3-12:

FIGS. 3-9 and 3-10 illustrate the effect of compounds 33 and 41according to the invention on triglyceride and cholesterol metabolism inApo E2/E2 transgenic mice. The animals were treated with the differentcompounds at a dose of 50 mg/kg/day for 8 days. FIGS. 5-1 and 5-2 showthe decrease in plasma triglycerides and cholesterol induced bycompounds 33 and 41.

FIGS. 3-11 and 3-12 show triglyceride and cholesterol distribution inlipoparticles evaluated by exclusion chromatography. A typicaldistribution of triglycerides and cholesterol is observed principally inlarge sized lipoparticles as well as a decrease in triglycerides andcholesterol in this lipoparticle subfraction under the effect ofcompounds 33 and 41.

Other aspects and advantages of the invention will become apparent inthe following examples, which are given for purposes of illustration andnot by way of limitation.

EXAMPLES

The inventive compounds were prepared according to the general methodsoutlined below.

Description of General Synthetic Methods of the Invention Synthesis of1,3-diphenyl prop-2-en-1-ones

General Method 1: Synthesis of 1,3-diphenylprop-2-en-1-ones in AcidicMedium

The ketone (1 eq) and the aldehyde (1 eq) were dissolved in ethanolsolution saturated with gaseous hydrochloric acid. The reaction wasstirred at room temperature for 6 hours and the solvent was theneliminated by vacuum evaporation. 1,3-diphenylprop-2-en-1-one waspurified by chromatography on silica gel.

General Method 2: Synthesis of 1,3-diphenylprop-2-en-1-ones in BasicMedium

The ketone (1 eq) and the aldehyde (1 eq) were dissolved in ahydroalcoholic solution of sodium hydroxide (20 eq). The mixture wasstirred at room temperature for 18 hours. The medium was acidified topH=2 with hydrochloric acid.

1,3-diphenylprop-2-en-1-one was obtained by precipitation orsolid/liquid extraction after evaporation of the reaction medium. It waspurified by silica gel chromatography or by recrystallization.

General Method 3: Synthesis of Substituted 1,3-diphenylprop-2-en-1-onesin the Presence of Sodium Ethylate

Sodium (1 eq) was dissolved in absolute ethanol. The ketone (1 eq) andthe aldehyde (1 eq) were added. The reaction mixture was stirred at roomtemperature for 12 hours and 2 N sodium hydroxide (5 eq) was then added.The mixture was kept at 100° C. for 12 hours. The reaction medium wasacidified by adding 6 N aqueous hydrochloric acid solution. The solventwas eliminated by vacuum evaporation. The residue was purified bychromatography on silica gel or by recrystallization.

O-Alkylation of phenols and S-alkylation of thiophenols

General Method 4:

The phenol (1 eq) was dissolved in acetonitrile. The halogenatedderivative (1 to 10 eq) and potassium carbonate (5 eq) were then added.The reaction medium was briskly stirred under reflux for approximately10 hours. The salts were eliminated by filtration, the solvent andexcess reagent were eliminated by vacuum evaporation, and the expectedproduct was purified by silica gel chromatography.

Acid Hydrolysis of Tertbutylic Esters: General Method 5:

The tertbutylic ester (1 eq) was dissolved in dichloromethane,trifluoroacetic acid (10 eq) was added, and the mixture was stirred atroom temperature for 12 hours. The resulting product was purified bychromatography on silica gel or by recrystallization.

Synthesis of Starting Materials Used to Synthesize the InventiveCompounds:

Starting Material 12′-Hydroxy-4′-(ethoxycarbonyldimethylmethoxy)acetophenone

This compound was synthesized from 2′,4′-dihydroxyacetophenone and ethylbromoisobutyrate (1 eq) according to general method 4 described earlier.It was purified by chromatography on silica gel (elution:cyclohexane/ethyl acetate 9:1).

1H NMR CDCl₃ δppm: 1.25 (t, J=7.17 Hz, 3H), 1.67 (s, 6H), 2.56 (s, 3H),4.24 (q, J=7.17 Hz, 2H), 6.27 (d, J=2.55 Hz, 1H), 6.37 (dd, J=2.55 Hz,J=8.72 Hz, 1H), 7.62 (d, J=8.72 Hz, 1H), 12.6 (signal, 1H).

Reference: U.S. Pat. No. 3,629,290 (1970), Fisons Pharmaceutical

Starting Material 2 3-chlorophenol acetate

3-chlorophenol was dissolved in dichloromethane. Triethylamine (1 eq)and acetic anhydride (2 eq) were added. The mixture was stirred at roomtemperature for 5 hours. Solvent was eliminated by vacuum evaporation.The evaporation residue was taken up in dichloromethane, dried onmagnesium sulfate and the solvent was eliminated by vacuum evaporation.Purification was carried out by chromatography on silica gel (elution:cyclohexane/ethyl acetate 95:5).

1H NMR CDCl₃ δppm: 2.29 (s, 3H), 6.99-7.33 (m, 4H)

Starting material 3 4′-Chloro-2′-hydroxyacetophenone

3-chlorophenyl acetate (starting material 2) was mixed with aluminiumchloride (3 eq). The mixture was heated at 200° C. for 1 hour. Thereaction medium was cooled. to room temperature then poured in ice. Theaqueous phase was extracted with methylene chloride which was dried onmagnesium sulfate then vacuum evaporated.

Purification was by silica gel chromatography (elution:cyclohexane/ethyl acetate 95:5).

1H NMR CDCl₃ δppm: 3.41 (s, 3H), 6.81 (dd, J=8.82 Hz, J=1.47 Hz, 1H),6.91 (d, J=1.47 Hz, 1H), 7.60 (d, 8.82 Hz, 1H), 12.33 (s, 1H)

Reference: Chen et al, J Chem Soc, 1958, 146-148.

Starting Material 4 4-Ethyloxycarbonyldimethylmethyloxybenzaldehyde

This compound was synthesized from 4-hydroxyabenzaldehyde and ethylbromoisobutyrate according to general method 4 described earlier.

Purification was carried out by silica gel chromatography (elution:cyclohexane/ethyl acetate 9:1).

1H NMR CDCl₃ δ ppm: 1.20 (t, J=6.96 Hz, 3H), 1.67 (s, 6H), 4.21 (q,J=6.96 Hz, 2H), 6.89 (d, J=8.91 Hz, 2H), 7.79 (d, J=8.94 Hz, 2H), 9.87(S, 1H).

Starting Material 53,5-dimethyloxy-4-ethyloxycarbonyldimethylmethyloxybenzaldehyde

This compound was synthesized from3,5-dimethyloxy-4-hydroxyabenzaldehyde and ethyl bromoisobutyrateaccording to general method 4 described earlier.

Purification was by silica gel chromatography (elution:cyclohexane/ethyl acetate 8:2).

1H NMR CDCl₃ δ ppm: 1.33 (t, J=7.29 Hz, 3H), 1.50 (s, 6H), 3.84 (s, 6H),4.27 (q, J=7.29 Hz, 2H), 7.08 (s, 2H), 9.86 (s, 1H)

Starting Material 63,5-dimethyl-4-ethyloxycarbonyldimethylmethyloxybenzaldehyde

This compound was synthesized from 3,5-dimethyl-4-hydroxyabenzaldehydeand ethyl bromoisobutyrate according to general method 4 describedearlier.

Purification was by silica gel chromatography (elution:cyclohexane/ethyl acetate 95:5).

1H NMR CDCl₃ δ ppm:1.37 (t, J=7.14 Hz, 3H), 1.50 (s, 6H), 2.29 (s, 6H),4.30 (q, J=7.14 Hz, 2H), 7.54 (s, 2H), 9.88 (s, 1H)

Starting Material 7 3-Ethyloxycarbonyldimethylmethyloxybenzaldehyde

This compound was synthesized from 3-hydroxybenzaldehyde and ethylbromoisobutyrate according to general method 4 described earlier.

Purification was by silica gel chromatography (elution:cyclohexane/ethyl acetate 9:1).

1H NMR CDCl₃ δppm: 1.24 (t, J=7.27 Hz, 3H), 1.62 (s, 6H), 4.25 (q,J=7.27 Hz, 2H), 7.11 (m, 1H), 7.31 (m, 1H), 7.40 (t, J=8.19 Hz, 1H),7.49 (m, 1H), 9.93 (s, 1H).

Starting Material 8 4-Ethyloxycarbonyldimethylmethyl thiobenzaldehyde

4-Methylthiobenzaldehyde (1 eq) was dissolved in methylene chloride andthe solution cooled to 0° C. Metachloroperbenzoic acid (1.5 eq) wasadded in small fractions. The reaction was followed by thin-layerchromatography. Additional metachloroperbenzoic acid was possibly addedso as to obtain total disappearance of the starting product. Theprecipitate was eliminated by filtration. Calcium hydroxide (1.5 eq) wasadded and the mixture was stirred for another 15 min. The solid waseliminated by filtration, the filtrate dried on magnesium sulfate andthe methylene chloride was then eliminated by vacuum evaporation.

The evaporation residue was taken up in acetic anhydride, then heatedunder reflux for 30 min and evaporated to dryness. The residue was takenup in methanol/triethylamine solution, stirred at room temperature for15 minutes, then the solvents were eliminated by vacuum evaporation. Theoily residue was taken up in a saturated aqueous ammonium chloridesolution then extracted with methylene chloride. The organic phase wasdried on magnesium sulfate and vacuum evaporated.

The resulting 4-mercaptobenzaldehyde intermediate was used withoutfurther purification. It was alkylated according to general method 4 toyield 4-ethyloxycarbonyldimethylmethylthiobenzaldehyde.

Purification was by silica gel chromatography (elution:cyclohexane/ethyl acetate 9:1).

1H NMR CDCl₃ δ ppm: 1.22 (t, J=7.46 Hz, 3H), 2.60 (s, 6H), 4.15 (q,J=7.46 Hz, 2H), 7.53 (d, J=8.38 Hz, 2H), 7.88 (d, J=8.39 Hz, 2H), 9.99(s, 1H)

Reference: Young N R, Gauthier J Y., Coombs W. (1984). TetrahedronLetters 25(17): 1753-1756.

Starting Material 9 4′-Ethyloxycarbonyldimethylmethyloxyacetophenone

This compound was synthesized from 4′-hydroxyacetophenone and ethylbromoisobutyrate according to general method 4 described earlier.

Purification was by silica gel chromatography (elution:cyclohexane/ethyl acetate 9:1).

1H NMR CDCl₃ δ ppm: 1.17 (t, J=5.64 Hz, 3H), 1.61 (s, 6H), 2.50 (s, 3H),4.18 (q, J=5.64 Hz, 2H), 6.78 (d, J=8.82 Hz, 2H), 7.83 (d, J=8.81 Hz,2H).

Starting Material 10 3-bromophenyl acetate

3-bromophenol was dissolved in dichloromethane. Triethylamine (1 eq) andacetic anhydride (2 eq) were added and the mixture was stirred at roomtemperature for 5 hours. The solvent was eliminated by vacuumevaporation. The evaporation residue was taken up in dichloromethanethen dried on magnesium sulfate. The solvent was eliminated by vacuumevaporation.

Purification was by silica gel chromatography (elution:cyclohexane/ethyl acetate 95:5).

1H NMR CDCl₃ δ ppm: 2.30 (s, 3H), 7.0-7.4 (m, 4H)

Starting Material 11 2′-hydroxy-4′-bromoacetophenone

3-bromophenyl acetate (starting material 10) was mixed with aluminiumchloride (3 eq), and the mixture was heated at 200° C. for 1 hour. Thereaction medium was cooled to room temperature then poured in ice. Theaqueous phase was extracted with methylene chloride which was dried onmagnesium sulfate.

Purification was by silica gel chromatography (elution:cyclohexane/ethyl acetate 95:5).

1H NMR CDCl₃ δ ppm: 2.59 (s, 3H), 7.01 (d, J=8.5 Hz, 1H), 7.13 (s, 1H),7.55 (d, J=8.5 Hz, 1H), 12.33 (s, 1H)

Starting Material 12 4′-Ethyloxycarbonyldimethylmethyloxyacetophenone

4′-methylthioacetophenone was dissolved in methylene chloride and thesolution cooled to 0° C. Metachloroperbenzoic acid (1.5 eq) was added insmall fractions. The reaction was followed by thin-layer chromatography.Additional metachloroperbenzoic acid was possibly added so as to obtaintotal disappearance of the starting product. The precipitate waseliminated by filtration. Calcium hydroxide (1.5 eq) was added and themixture was stirred for another 15 min. The solid was eliminated byfiltration, the filtrate dried on magnesium sulfate and the methylenechloride was then eliminated by vacuum evaporation.

The evaporation residue was taken up in acetic anhydride, then heatedunder reflux for 30 min and evaporated to dryness. The residue was takenup in methanol/triethylamine solution, stirred at room temperature for15 minutes, then the solvents were eliminated by vacuum evaporation. Theoily residue was taken up in a saturated aqueous ammonium chloridesolution then extracted with methylene chloride. The organic phase wasdried on magnesium sulfate then vacuum evaporated.

The resulting 4-mercaptoacetophenone intermediate was used withoutfurther purification. It was alkylated according to general method 4 toyield 4-ethyloxycarbonyldimethylmethylthioacetophenone.

Purification was by silica gel chromatography (elution:cyclohexane/ethyl acetate 9:1).

Reference: Young N R, Gauthier J Y., Coombs w (1984). TetrahedronLetters 25(17): 1753-1756.

1H NMR CDCl₃ δ ppm: 1.21 (t, J=7.32 Hz, 3H), 1.51 (s, 6H), 2.59 (s, 3H),4.12 (q, J=7.32 Hz, 2H), 7.51 (d, J=8.40 Hz, 2H), 7.79 (d, J=8.40 Hz,2H)

Synthesis of Intermediate Compounds Used to Synthesize the InventiveCompounds: Intermediate Compound 11-14-chlorophenyl]-3-[3,5-dimethyl-4-hydroxyphenyl]prop-2-en-1-one

This compound was synthesized from 4-chloroacetophenone and3,5-dimethyl-4-hydroxybenzaldehyde according to general method 1described earlier.

Purification was by chromatography on silica gel (elution:cyclohexane/ethyl acetate 95:5).

1H NMR CDCl₃ δ ppm: 2.30 (s, 6H), 7.32 (s, 2H), 7.34 (d, J=15.25 Hz,1H), 7.47 (d, J=8.86 Hz, 2H), 7.75 (d, J=15.26 Hz, 1H), 7.97 (d, J=8.86Hz, 2H).

Intermediate Compound 21-[4-methylthiophenyl]-3-[3,5-dimethyl-4-hydroxyphenyl]prop-2-en-1-one

This compound was synthesized from 4′-methylthioacetophenone and3,5-dimethyl-4-hydroxybenzaldehyde according to general method 1described earlier.

Purification was by chromatography on silica gel (elution:cyclohexane/ethyl acetate 8:2).

1H NMR DMSO δ ppm: 2.22 (s, 6H), 2.54 (s, 3H), 7.36 (d, J=8.20 Hz, 2H),7.48 (s, 2H), 7.62 (d, J=15.7 Hz, 1H), 7.74 (d, J=15.7 Hz, 1H), 8.10 (d,J=8.20 Hz, 2H), 8.92 (s, 1H)

Intermediate Compound 31-[2-methoxyphenyl]-3-[3,5-dimethyl-4-hydroxyphenyl]prop-2-en-1-one

This compound was synthesized from 2′-methoxyacetophenone and3,5-dimethyl-4-hydroxybenzaldehyde according to general method 1described earlier.

Purification was by chromatography on silica gel (elution:cyclohexane/ethyl acetate 8:2).

1H NMR DMSO δ ppm: 2.39 (s, 6H), 2.22 (s, 6H), 7.58 (s, 2H), 7.67-7.62(m, 3H), 7.82 (d, J=15.5 Hz, 1H), 8.17 (d, 1H), 12.96 (s, 1H)

Intermediate Compound 41-[4-hexyloxyphenyl]-3-[3,5-dimethyl-4-hydroxyphenyl]prop-2-en-1-one

This compound was synthesized from 4-hexyloxyacetophenone and3,5-dimethyl-4-hydroxybenzaldehyde according to general method 1described earlier.

The expected compound precipitated in the reaction medium; it was driedand used without further purification for the following reaction.

1H NMR DMSO δ ppm: 0.88 (m, 3H), 1.28-1.43 (m, 6H), 1.72 (m, 2H), 2.21(s, 6H), 4.05 (t, J=6.42 Hz, 2H), 7.40 (d, J=8.43 Hz, 2H), 7.48 (s, 2H),7.57 (d, J=15.24 Hz, 1H), 7.72 (d, J=15.24 Hz, 1H), 8.12 (d, J=8.43 Hz,2H), 8.89 (s, 1H)

Intermediate Compound 51-[2-hydroxy-4-chlorophenyl]-3-[3,5-dimethyl-4-hydroxyphenyl]prop-2-en-1-one

This compound was synthesized from 4′-chloro-2′-hydroxyacetophenone(starting material 3) and 3,5-dimethyl-4-hydroxybenzaldehyde accordingto general method 1 described earlier.

Purification was by chromatography on silica gel (toluene: 10).

1H NMR DMSO δppm: 2.21 (s, 6H), 7.1 (m, 2H), 7.55 (s, 2H), 7.72 (d,J=15.4 Hz, 1H), 7.80 (d, J=15.4 Hz, 1H), 8.25 (d, J=9.0 Hz, 1H), 9.09(s, 1H), 13.04 (s, 1H)

Intermediate Compound 62-(3,5-dimethyl-4-hydroxyphenyl)-7-chloro-4H-1-benzopyran-4-one

This compound was synthesized from1-[2-hydroxy-4-chlorophenyl]-3-[3,5-dimethyl-4-hydroxyphenyl]prop-2-en-1-one(intermediate compound 5) according to the following method:

1-[2-hydroxy-4-chlorophenyl]-3-[3,5-dimethyl-4-hydroxyphenyl]prop-2-en-1-onewas dissolved in dimethylsulfoxide, an iodine crystal was added, and themixture was kept under reflux for 10 min.

The reaction medium was brought to room temperature, hydrolyzed. Theprecipitate was dried, rinsed with sodium thiosulfate solution then withwater.

Purification was by dissolution in methylene chloride and precipitationby addition of heptane.

1H NMR DMSO δppm: 2.25 (s, 6H), 6.87 (s, 1H), 7.51 (d, J=8.55 Hz, 1H),7.73 (s, 2H), 7.98 (m, 2H)

Reference: Doshi A G, S. P., Ghiya B J (1986). Indian J Chem Sect B 25:759.

Intermediate Compound 71-[2-methyloxy-4-chlorophenyl]-3-[3,5-dimethyl-4-hydroxydimethylmethyloxyphenyl]prop-2-en-1-one

This compound was synthesized from 4′-chloro-2′-methoxyacetophenone and3,5-dimethyl-4-hydroxybenzaldehyde according to general method 1described earlier.

Purification was by chromatography on silica gel (elution:cyclohexane/ethyl acetate 85:15).

1H NMR DMSO δ ppm: 2.21 (s, 6H), 3.90 (s, 3H), 7.12 (m, 1H), 7.23 (d,J=15.5 Hz, 1H), 7.29 (s, J=1.80 Hz, 1H), 7.38 (d, J=15.5 Hz, 1H), 7.41(s, 2H), 7.48 (d, J=7.98 Hz, 1H)

Intermediate Compound 81-[4-bromophenyl]-3-[3,5-dimethyl-4-hydroxyphenyl]prop-2-en-1-one

This compound was synthesized from 4′-bromoacetophenone and3,5-dimethyl-4-hydroxybenzaldehyde according to general method 1described earlier.

Purification was by chromatography on silica gel (elution:cyclohexane/ethyl acetate 85:15).

1H NMR DMSO δ ppm: 2,30 (s, 6H), 7.32 (s, 2H), 7.56-7.66 (m, 3H), 7.75(d, J=15.27 Hz, 1H), 7.90 (d, J=8.70 Hz, 2H), 9.82 (s, 1H)

Intermediate Compound 91-[4-heptylphenyl]-3-[3,5-dimethyl-4-hydroxyphenyl]prop-2-en-1-one

This compound was synthesized from 4′-heptylacetophenone and3,5-dimethyl-4-hydroxybenzaldehyde according to general method 1described earlier.

Purification was by chromatography on silica gel (elution:cyclohexane/ethyl acetate 85:15).

1H NMR DMSO δ ppm: 0.84 (m, 3H), 1.25 (m, 8H), 1.60 (m, 2H), 2.21 (s,6H), 2.65 (t, J=7.50 Hz, 2H), 7.35 (d, J=8.02 Hz, 2H), 7.48 (s, 2H),7.60 (d, J=15.48 Hz, 1H), 7.71 (d, J=15.48 Hz, 1H), 8.05 (d, J=8.02 Hz,2H), 8.92 (s, 1H)

Synthesis of the Inventive Compounds Compound 11-[2-hydroxy-4-ethoxycarbonyldimethylmethyloxyphenyl]-3-[3,5-ditertbutyl-4-hydroxyphenyl]prop-2-en-1-one

This compound was synthesized from2′-hydroxy-4′-(ethoxycarbonyldimethylmethoxy)acetophenone (startingmaterial 1) and 3,5-ditertbutyl-4-hydroxybenzaldehyde according togeneral method 1 described earlier.

Purification was by chromatography on silica gel (elution:cyclohexane/ethyl acetate 9:1).

1H NMR CDCl₃ δ ppm: 1.25 (t, J=7.11 Hz, 3H), 1.45 (s, 18H), 1.70 (s,6H), 4.26 (q, J=7.11 Hz, 2H), 5.63 (s, 1H), 6.33 (d, J=2.37 Hz, 1H),6.42 (dd, J=8.8 Hz, J=2.37 Hz, 1H), 7.41 (d, J=15.39 Hz, 1H), 7.5 (s,2H), 7.83 (d, J=8.8 Hz, 1H), 7.88 (J=15.39 Hz, 1H), 13.5 (s, 1H)

Compound 21-[2-hydroxy-4-carboxydimethylmethyloxyphenyl]-3-[3,5-ditertbutyl-4-hydroxyphenyl]prop-2-en-1-one

This compound was synthesized from1-[2-hydroxy-4-ethoxycarbonyldimethylmethyloxyphenyl]-3-[3,5-ditertbutyl-4-hydroxyphenyl]prop-2-en-1-one(compound 1) according to the following method:

The ester was dissolved in ethanol, an aqueous 1N sodium hydroxidesolution (5 eq) was added, and the mixture was kept under reflux for 10hours. The medium was acidified by addition of 12 N hydrochloric acidthen extracted with ethyl acetate. The organic phase was dried onmagnesium sulfate then vacuum evaporated.

Purification was by preparative HPLC (reverse phase RP18, Licrospher 12μm, elution: water-methanol-trifluoroacetic acid: 22:78:0.1).

1H NMR CDCl₃ δ ppm: 1.49 (s, 18H), 1.73 (s, 6H), 5.62 (s, 1H), 6.44 (d,J=15.5 Hz, 1H), 7.01 (m, 2H), 7.57 (t, 1H), 7.81 (d, J=15.5 Hz, 1H),7.87 (d, 2H), 7.93 (d, 1H), 8.26(d, 1H)

MS (ES-MS): 453.2 (M−1)

Compound 31-[2-hydroxy-4-chlorophenyl]-3-[4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one

This compound was synthesized from 2′-hydroxy-4′-chloroacetophenone and4-ethyloxycarbonyldimethylmethyloxybenzaldehyde (starting material 9)according to general method 2 described earlier.

Purification was by chromatography on silica gel (elution:cyclohexane/ethyl acetate 9:1).

1H NMR DMSO δ ppm: 1.58 (s, 6H), 6.87 (d, J=8.54 Hz, 2H), 7.05 (dd,J=8.54 Hz, 1.83 Hz, 1H), 7.09 (d, J=1.2 Hz, 1H), 7.90-7.80 (m, 4H), 8.25(m, 8.52 Hz, 1H), 12.84 (s, 1H), 13.26 (s, 1H)

MS (ES-MS): 359.0 (M−1)

Compound 41-[2-hydroxyphenyl]-3-[4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one

This compound was synthesized from 2′-hydroxyacetophenone and4-ethyloxycarbonyldimethylmethyloxybenzaldehyde (starting material 4)according to general method 2 described earlier.

Purification was carried out by chromatography on silica gel (elution:cyclohexane/ethyl acetate 9:1).

1H NMR DMSO δ ppm: 1.58 (s, 6H), 6.88 (d, 2H), 7.01 (m, 2H), 7.57 (t,1H), 7.81 (d, J=15.5 Hz, 1H), 7.87 (d, 2H), 7.93 (d, J=15.5 Hz, 1H),8.26 (d, 1H), 12.69 (s, 1H)

MS (ES-MS): 325.1 (M−1)

Compound 51-[2-hydroxyphenyl]-3-[3,5-dimethoxy-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one

This compound was synthesized from 2′-hydroxyacetophenone and3,5-dimethyloxy-4-ethyloxycarbonyldimethylmethyloxybenzaldehyde(starting material 5) according to general method 2 described earlier.

Purification was carried out by chromatography on silica gel (elution:cyclohexane/ethyl acetate 9:1).

1H NMR DMSO δ ppm: 1.35 (s, 6H), 3.80 (s, 6H), 7.00-7.03 (m, 2H), 7.25(s, 2H), 7.59 (t, 1H, J=8.07 Hz, 1H), 7.81 (d, J=15.5 Hz, 1H), 8.00 (d,J=15.5 Hz, 1H), 8.31 (d, J=8.07 Hz, 1H), 12.36 (s, 1H), 12.69 (s, 1H)

MS (ES-MS): 385.3 (M−1)

Compound 61-[2-hydroxy-4-chlorophenyl]-3-[3,5-dimethoxy-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one

This compound was synthesized from 2′-hydroxy-4′-chloroacetophenone(starting material 3) and3,5-dimethyloxy-4-ethyloxycarbonyldimethylmethyloxybenzaldehyde(starting material 5) according to general method 2 described earlier.

Purification was carried out by chromatography on silica gel (elution:cyclohexane/ethyl acetate 9:1).

1H NMR DMSO δ ppm: 1.34 (s, 6H), 3.80 (s, 6H), 7.08 (dd, J=1.77 Hz, 1H),7.12 (d, J=1.77 Hz,1H), 7.24 (s, 2H), 7.79 (d, J=15.4 Hz, 1H), 7.93 (d,J=15.4 Hz, 1H), 8.27 (d, J=8.3 Hz, 1H), 12.36 (s, 1H), 12.69 (s, 1H)

MS (ES-MS): 419.0 (M−1)

Compound 71-[2-hydroxy-4-chlorophenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one

This compound was synthesized from 2′-hydroxy-4′-chloroacetophenone(starting material 3) and3,5-dimethyl-4-ethyloxycarbonyldimethylmethyloxybenzaldehyde (startingmaterial 6) according to general method 2 described earlier.

Purification was carried out by chromatography on silica gel (elution:cyclohexane/ethyl acetate 9:1).

1H NMR DMSO δ ppm: 1.39 (s, 6H), 2.22 (s, 6H), 7.07 (m, 1H), 7.12 (d,J=2.07 Hz, 1H), 7.61 (s, 2H), 7.74 (d, J=15.5 Hz, 1H), 7.87 (d, J=15.5Hz, 1H), 8.26 (d, 1H), 12.76 (s, 1H)

MS (ES-MS): 387.1 (M−1)

Compound 81-[2-hydroxy-4-carboxydimethylmethyloxyphenyl]-3-[3,5-dibromo-4-hydroxyphenyl]prop-2-en-1-one

This compound was synthesized from 2′-hydroxy-4′-ethyloxycarbonyldimethylmethyloxyacetophenone (starting material 1) and3,5-dibromo-4-hydroxybenzaldehyde according to general method 2described earlier.

Purification was carried out by chromatography on silica gel (elution:cyclohexane/ethyl acetate 9:1).

1H NMR CDCl₃ δ ppm: 1.60 (s, 6H), 6.24 (d, J=2.47 Hz, 1H), 6.43 (dd,J=2.47 Hz, J=8.52 Hz, 1H), 7.70 (d, J=15.5 Hz, 1H), 7.96 (d, J=15.5 Hz,1H), 8.22 (s, 2H), 8.34 (d, J=9.16 Hz, 1H), 13.34 (s, 1H)

MS (ES-MS): 498.6 (M−1)

Compound 91-[2-hydroxyphenyl]-3-[3-carboxydimethylmethyloxyphenyl]prop-2-en-1-one

This compound was synthesized from 2′-hydroxyacetophenone and3-ethyloxycarbonyldimethylmethyloxybenzaldehyde (starting material 7)according to general method 2 described earlier.

Purification was carried out by chromatography on silica gel (elution:cyclohexane/ethyl acetate 9:1).

1H NMR DMSO δ ppm: 1.56 (s, 6H), 6.91 (dd, J=8.01 Hz, J=2.47 Hz, 1H),7.03-6.99 (m, 2H), 7.41-7.36 (m, 2H), 7.60-7.52 (m, 2H), 7.77 (d, J=15.5Hz, 1H), 8.00 (d, J=15.5 Hz, 1H), 8.31 (dd, J=8.63 Hz, J=1.85 Hz, 1H),12.47 (s, 1H), 13.17 (s, 1H)

MS (ES-MS): 325.8(M−1)

Compound 101-[2-hydroxy-4-carboxydimethylmethyloxyphenyl]-3-[3-hydroxyphenyl]prop-2-en-1-one

This compound was synthesized from 2′-hydroxy-4′-ethyloxycarbonyldimethylmethyloxyacetophenone (starting material 1) and3-hydroxybenzaldehyde according to general method 2 described earlier.

Purification was carried out by chromatography on silica gel (elution:cyclohexane/ethyl acetate 9:1).

1H NMR DMSO δ ppm: 1.60 (s, 6H), 6.25 (d, J=2.47 Hz, 1H), 6.43 (dd,J=2.47 Hz, 9.09 Hz, 1H), 6.89 (m, 1H), 7.35-7.24 (m, 3H), 7.73 (d, 1H),7.92 (d, J=15.5 Hz, 1H), 8.27 (d, J=15.5 Hz, 1H), 13.21 (s, 1H), 13.39(s, 1H).

MS (ES-MS): 341(M−1)

Compound 111-[2-hydroxyphenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one

This compound was synthesized from 2′-hydroxyacetophenone and3,5-dimethyl-4-ethyloxycarbonyldimethylmethyloxybenzaldehyde (startingmaterial 6) according to general method 2 described earlier.

Purification was carried out by chromatography on silica gel (elution:cyclohexane/ethyl acetate: 9/1) followed by preparative HPLC (reversephase RP18, Licrospher 12 μm, elution: water/methanol/trifluoroaceticacid: 22/78/0.1).

1H NMR DMSO δ ppm: 1.57 (s, 6H), 2.31 (s, 6H), 6.96 (t, J=8.17 Hz, 1H),7.04 (d, J=8.72 Hz, 1H), 7.35 (s, 2H), 7.49 (t, J=8.2 Hz, 1H), 7.58 (d,J=15.8 Hz,1H), 7.84 (d, J=15.8 Hz, 1H), 7.94 (d, J=8.7 Hz, 1H), 12.87(s, 1H)

MS (ES-MS): 353.1 (M−1)

Compound 121-[2-hydroxy-4-carboxydimethylmethyloxyphenyl]-3-[4-methylthiophenyl]prop-2-en-1-one

This compound was synthesized from2′-hydroxy-4′-ethyloxycarbonyldimethylmethyloxyacetophenone (startingmaterial 1) and 4-methylthiobenzaldehyde according to general method 2described earlier. Purification was by chromatography on silica gel(elution: cyclohexane/ethyl acetate 9/1) followed by preparative HPLC(reverse phase RP18, Licrospher 12 μm, elution:water/methanol/trifluoroacetic acid: 22/78/0.3).

1H NMR DMSO δ ppm: 1.60 (s, 6H), 2.54 (s, 3H), 6.25 (d, 1H), 6.43 (dd,J=2.47 Hz, 1H), 7.33 (d, J=8.56 Hz, 2H), 7.8 (d, 15.5 Hz, 1H), 7.86 (d,J=8.56 Hz, 2H), 7.98 (d, J=15.5 Hz, 1H), 8.29 (d, J=9.1 Hz, 1H), 13.34(s, 1H)

MS (ES-MS): 373.1 (M−1)

Compound 131-[2,4-dihydroxyphenyl-3-[4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one

This compound was synthesized from 2′,4′-dihydroxyacetophenone and4-ethoxycarbonyldimethylmethyloxybenzaldehyde (starting material 4)according to general method 2 described earlier.

Purification was by chromatography on silica gel (elution:cyclohexane/ethyl acetate 9/1) followed by preparative HPLC (reversephase RP18, Licrospher 12 μm, elution: water/methanol/trifluoroaceticacid: 34/66/0.1).

1H NMR DMSO δ ppm: 1.57 (s, 6H), 6.29 (d, J=2.16 Hz, 1H), 6.41 (dd,J=9.18 Hz, J=2.16 Hz, 1H), 6.86 (d, J=8.64 Hz, 2H), 7.75 (d, J=15.67 Hz,1H), 7.83-7.88 (m, 3H), 8.19 (d, J=9.18 Hz, 1H), 10.74 (s, 1H), 13.53(s, 1H)

MS (maldi-Tof): 343.1(M+1)

Compound 14 1-[2-hydroxyphenyl]-3-[4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one

This compound was synthesized from 4′-hydroxyacetophenone and4-ethoxycarbonyldimethylmethyloxybenzaldehyde (starting material 4)according to general method 2 described earlier.

Purification was carried out by chromatography on silica gel (elution:cyclohexane/ethyl acetate 9:1) followed by preparative HPLC (reversephase RP18, Licrospher 12 μm, elution: water/methanol/trifluoroaceticacid: 34/66/0.1).

1H NMR DMSO δ ppm: 1.56 (s, 6H), 6.85 (d, J=8.63 Hz, 2H), 6.90 (d,J=9.21 Hz, 2H), 7.63 (d, J=15.54 Hz, 1H), 7.78 (m, 3H), 8.05 (d, J=8.61Hz, 2H), 10.40 (s, 1H), 13.22 (s, 1H)

MS (maldi-Tof): 327.1(M+1)

Compound 151-[4-chlorophenyl]-3-[3,5-dimethyl-4-isopronyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one

This compound was synthesized from1-[4-chlorophenyl]-3-[3,5-dimethyl-4-hydroxyphenyl]prop-2-en-1-one(intermediate compound 1) and isopropyl bromoisobutyrate according togeneral method 4 described earlier.

Purification was carried out by chromatography on silica gel (elution:cyclohexane/ethyl acetate 9/1).

1H NMR DMSO δ ppm: 1.25 (d, J=6.06 Hz, 6H), 1.39 (s, 6H), 5.00 (sept,J=6.06 Hz, 1H), 7.57 (s, 2H), 7.62 (d, J=8.40 Hz, 2H), 7.64 (d, J=15.8Hz, 1H), 7.81 (d, J=15.8 Hz, 1H), 8.16 (d, J=8.40 Hz, 2H).

MS (Maldi-Tof): 415.1(M+1)

Compound 161-[4-chlorophenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one

This compound was synthesized from1-[4-chlorophenyl]-3-[3,5-dimethyl-4-hydroxyphenyl]prop-2-en-1-one(intermediate compound 1) and tertbutyl bromoisobutyrate according togeneral method 4 described earlier.

Purification was by chromatography on silica gel (elution:cyclohexane/ethyl acetate 9/1).

Compound 171-[4-chlorophenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one

This compound was synthesized from1-[4-chlorophenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one(compound 16) according to general method 5 described earlier.

Purification was carried out by chromatography on silica gel (elution:dichloromethane/methanol 98/2)

1H NMR DMSO δ ppm: 1.39 (s, 6H), 2.22 (s, 6H), 7.58 (s, 2H), 7.67-7.62(m, 3H), 7.82 (d, J=15.5 Hz, 1H), 8.17 (d, 1H), 12.96 (s, 1H)

MS (Maldi-Tof): 373.3(M+1)

Compound 181-[2-hydroxy-4-carboxydimethylmethyloxyphenyl]-3-[4-chlorophenyl]prop-2-en-1-one

This compound was synthesized from2′-hydroxy-4′-ethyloxycarbonyldimethylmethyloxyacetophenone (startingmaterial 1) and 4-chlorobenzaldehyde according to general method 2described earlier.

Purification was by chromatography on silica gel (elution:cyclohexane/ethyl acetate 9:1) followed by preparative HPLC (reversephase RP18, Licrospher 12 μm, elution: water/methanol/trifluoroaceticacid: 22/78/0.1).

1H NMR DMSO δ ppm: 1.60 (s, 6H), 6.25 (d, J=2.47 Hz, 1H), 6.45 (dd,J=2.47 Hz, J=9.12 Hz, 1H), 6.55 (d, J=8.55 Hz, 2H), 7.82 (d, J=15.54 Hz,1H), 7.97 (d, J=8.55 Hz, 2H), 8.03 (d, J=15.54 Hz, 1H), 8.29 (d, J=9.12Hz, 1H), 13.20 (s, 1H), 13.39 (s, 1H)

MS (ES-MS): 359.0 (M−1)

Compound 19 1-[2-hydroxyphenyl-3-[4-carboxydimethylmethylthiophenyl]prop-2-en-1-one

This compound was synthesized from 2′-hydroxyacetophenone andethyloxycarbonyldimethylmethylthiobenzaldehyde (starting material 8)according to general method 2 described earlier.

Purification was by chromatography on silica gel (elution:dichloromethane/methanol 95/5) followed by preparative HPLC (reversephase RP18, Licrospher 12 μm, elution: water/methanol/trifluoroaceticacid: 22/78/0.1).

1H NMR DMSO δ ppm: 1.44 (s, 6H), 6.99-7.05 (m, 1H), 7.52 (d, J=8.1 Hz,2H), 7.58 (m, 1H), 7.83 (d, J=15.5 Hz, 1H), 7.92 (d, J=8.1 Hz, 1H), 8.09(d, J=15.5 Hz, 1H), 8.26 (dd, J=1.62, J=8.6 Hz, 1H), 12.47 (s, 1H),12.78 (s, 1H) MS (Maldi-Tof): 242.9 (M+1)

Compound 201-[4-chloro-2-hydroxyphenyl]-3-[4-carboxydimethylmethylthiophenyl]prop-2-en-1-one

This compound was synthesized from 4′-chloro-2′-hydroxyacetophenone(starting material 3) and4-ethyloxycarbonyldimethylmethylthiobenzaldehyde (starting material 8)according to general method 2 described earlier.

Purification was by chromatography on silica gel (elution:dichloromethane/methanol 95:5) followed by preparative HPLC (reversephase RP18, Licrospher 12 μm, elution: water/methanol/trifluoroaceticacid: 22/78/0.1).

1H NMR DMSO δ ppm: 1.43 (s, 6H), 7.05 (dd, J=1.7 Hz, J=8.46 Hz, 1H),7.11 (d, J=2.25 Hz, 1H), 7.51 (d, J=7.92 Hz, 2H), 7.82 (d, J=15.8 Hz,1H), 7.89 (d, J=7.9 Hz, 2H), 8.05 (d, J=15.2 Hz, 1H), 8.23 (d, J=8.46Hz, 1H), 12.57 (s, 1H), 12.78 (s, 1H).

MS (Maldi-Tof): 377.0(M−1)

Compound 211-[4-carboxydimethylmethyloxyphenyl]-3-[3,5-dimethyl4-hydroxyphenyl]prop-2-en-1-one

This compound was synthesized from 4-ethyloxycarbonyldimethylmethyloxyacetophenone (starting material 9) and3,5-dimethyl-4-hydroxybenzaldehyde according to general method 2described earlier.

Purification was by chromatography on silica gel (elution:dichloromethane/methanol 95:5) followed by preparative HPLC (reversephase RP18, Licrospher 12 μm, elution: water/methanol/trifluoroaceticacid: 22/78/0.1).

1H NMR DMSO δ ppm: 1.60 (s, 6H), 2.21 (s, 6H), 6.91 (d, J=9.09 Hz, 2H),7.48 (s, 2H), 7.57 (d, J=15.12 Hz, 1H), 7.70 (d, J=15.63 Hz, 1H), 8.09(d, J=9.06 Hz, 2H), 8.9 (s, 1H), 13.29 (s, 1H)

MS (Maldi-Tof): 355.2 (M+1)

Compound 22 1-[4-methylthiophenyl[-3-[4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one

This compound was synthesized from 4′-methylthioacetophenone (startingmaterial 12) and 4-ethyloxycarbonyldimethylmethyloxybenzaldehyde(starting material 9) according to general method 2 described earlier.

Purification was by chromatography on silica gel (elution:dichloromethane/methanol 95:5) followed by preparative HPLC (reversephase RP18, Licrospher 12 μm, elution: water/methanol/trifluoroaceticacid: 22/78/0.1).

1H NMR DMSO δ ppm: 1.57 (s, 6H), 2.57 (s, 3H), 6.86 (d, J=8.94 Hz, 2H),7.41 (d, J=8.40 Hz, 2H), 7.69 (d, J=15.2 Hz, 1H), 7.84-7.78 (m, 3H),8.09 (d, J=8.4 Hz, 2H), 13.21 (s, 1H)

MS (Maldi-Tof): 357.2 (M−1)

Compound 231-[4-carboxydimethylmethyloxyphenyl]-3-[4-chlorophenyl]prop-2-en-1-one

This compound was synthesized from 4-ethyloxycarbonyldimethylmethyloxyacetophenone (starting material 9) and4-chlorobenzaldehyde according to general method 3 described earlier.

Purification was by chromatography on silica gel (elution:dichloromethane/methanol 95:5) followed by preparative HPLC (reversephase RP18, Licrospher 12 μm, elution: water/methanol/trifluoroaceticacid: 22/78/0.1).

1H NMR DMSO δ ppm: 1.72 (s, 6H), 6.97 (d, J=8.61 Hz, 2H), 7.39 (d,J=8.25 Hz, 2H), 7.50 (d, J=15.72 Hz, 1H), 7.57 (d, J=8.61 Hz, 2H), 7.77(d, J=15.72 Hz, 1H), 7.99 (d, J=8.61 Hz, 2H), 13.30 (s, 1H)

MS (Maldi-Tof): 345.1(M+1)

Compound 241-[4-carboxydimethylmethylthiophenyl]-3-[4-methylthiophenyl]prop-2-en-1-one

This compound was synthesized from 4-ethyloxycarbonyldimethylmethylthioacetophenone (starting material 12) and4-methylthiobenzaldehyde according to general method 3 describedearlier.

Purification was by chromatography on silica gel (elution:dichloromethane/methanol 95:5) followed by preparative HPLC (reversephase RP18, Licrospher 12 μm, elution: water/methanol/trifluoroaceticacid: 22/78/0.1).

1H NMR DMSO δ ppm: 1.46 (s, 6H), 2.54 (s, 3H), 7.33 (d, J=8.61 Hz, 2H),7.59 (d, J=8.10 Hz, 2H), 7.73 (d, J=15.66 Hz, 1H), 7.85 (d, J=8.10 Hz,2H), 7.92 (d, J=15.66 Hz, 1H), 8.13 (d, 8.10 Hz, 2H), 12.85 (s, 1H)

MS (Maldi-Tof): 373.1 (M+1)

Compound 251-[2-hydroxy-4-bromophenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one

This compound was synthesized from 4′-bromo-2′-hydroxyacetophenone(starting material 11) and3,5-dimethyl-4-ethyloxycarbonyldimethyloxybenzaldehyde (startingmaterial 6) according to general method 2 described earlier.

Purification was by chromatography on silica gel (elution:dichloromethane/methanol 95:5) followed by preparative HPLC (reversephase RP18, Licrospher 12 μm, elution: water/methanol/trifluoroaceticacid: 22/78/0.1).

1H NMR DMSO δ ppm: 1.39 (s, 6H), 2.22 (s, 6H), 7.20 (dd, J=2.16, J=8.55Hz, 1H), 7.25 (d, J=1.59 Hz, 1H), 7.60 (s, 2H), 7.73 (d, J=15.51 Hz,1H), 7.86 (d, J=15.51 Hz, 1H), 8.16 (d, J=8.58 Hz, 1H), 12.70 (s, 1H),13.30 (s, 1H)

MS (ES-MS): 432.9 (M−1)

Compound 261-[4-carboxydimethylmethyloxyphenyl]-3-[4-methylthiophenyl]prop-2-en-1-one

This compound was synthesized from4′-ethyloxycarbonyldimethylmethyloxyacetophenone (starting material 9)and 4-methylthiobenzaldehyde according to general method 2 describedearlier.

Purification was by chromatography on silica gel (elution:dichloromethane/methanol 95:5) followed by preparative HPLC (reversephase RP18, Licrospher 12 μm, elution: water/methanol/trifluoroaceticacid: 22/78/0.1).

1H NMR DMSO δ ppm: 1.60 (s, 6H), 2.53 (s, 3H), 6.93 (d, J=9.00 Hz, 2H),7.32(d, J=8.49 Hz, 2H), 7.68 (d, J=15.51 Hz, 1H), 7.82 (d, J=8.52 Hz,2H), 7.89 (d, J=15.51 Hz, 1H), 8.13 (d, 9.00 Hz, 2H), 13.30 (s, 1H)

MS (Maldi-Tof): 355.0(M+1)

Compound 271-[4-methylthiophenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one

This compound was synthesized from1-[4-methylthiophenyl]-3-[3,5-dimethyl-4-hydroxyphenyl]prop-2-en-1-one(intermediate compound 2) and tertbutyl bromoisobutyrate according togeneral method 4 described earlier.

Purification was by chromatography on silica gel (elution:cyclohexane/ethyl acetate 8/2).

Compound 281-[4-methylthiophenyl]-3-[3,5-dimethyl-4-isopropyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one

This compound was synthesized from1-[4-methylthiophenyl]-3-[3,5-dimethyl-4-hydroxyphenyl]prop-2-en-1-one(intermediate compound 2) and isopropyl bromoisobutyrate according togeneral method 4 described earlier. Purification was by chromatographyon silica gel (elution: cyclohexane/ethyl acetate 9/1).

1H NMR DMSO δ ppm: 1.25 (d, J=6.18 Hz, 6H), 1.39 (s, 6H), 2.18 (s, 6H),2.57 (s, 3H), 4.99 (sept, J=6.18 Hz, 1H), 7.40 (d, J=8.28 Hz, 2H), 7.58(s, 2H), 7.62 (d, J=15.5 Hz, 1H), 7.82 (d, J=15.5 Hz, 1H), 8.10 (d,=8.28 Hz, 2H), 12.97 (s, 1H)

MS (Maldi-Tof): 427.1 (M+1)

Compound 291-[4-methylthiophenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one

This compound was synthesized from1-[4-methylthiophenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one(compound 28) according to general method 5 described earlier.

Purification was by chromatography on silica gel (elution:dichloromethane /methanol 98/2).

1H NMR DMSO δ ppm: 1.39 (s, 6H), 2.22 (s, 6H), 2.57 (s, 3H), 7.40 (d,J=8.55 Hz, 2H), 7.57 (s, 2H), 7.62 (d, J=15.5 Hz, 1H), 7.83 (d, J=15.5Hz, 1H), 8.10 (d, J=8.55 Hz, 2H), 12.97 (s, 1H)

MS (ES-MS): 383.3(M−1)

Compound 301-[2-methoxyphenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one

This compound was synthesized from1-[2-methoxyphenyl]-3-[3,5-dimethy-l-4-hydroxyphenyl]prop-2-en-1-one(intermediate compound 3) and tertbutyl bromoisobutyrate according togeneral method 4 described earlier.

Purification was by chromatography on silica gel (elution:cyclohexane/ethyl acetate 9:1).

Compound 311-[2-methoxyphenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one

This compound was synthesized from1-[2-methoxyphenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one(compound 30) according to general method 5 described earlier.

Purification was by chromatography on silica gel (elution:dichloromethane/methanol 98/2).

1H NMR DMSO δ ppm: 1.38 (s, 6H), 2.19 (s, 6H), 3.93 (s, 3H), 7.05 (m,1H), 7.20 (d, J=8.31 Hz, 1H), 7.25 (d, J=15.5 Hz, 1H), 7.37 (d, J=15.5Hz, 1H), 7.39 (s, 2H), 7.46 (d, J=7.2 Hz, 1H), 7.53 (m, 1H), 12.93 (s,1H)

MS (ES-MS): 367.1(M−1)

Compound 32 1-[4-hexyloxyphenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one

This compound was synthesized from1-[4-hexyloxyphenyl]-3-[3,5-dimethyl-4-hydroxyphenyl]prop-2-en-1-one(intermediate compound 4) and tertbutyl bromoisobutyrate according togeneral method 4 described earlier.

Purification was by chromatography on silica gel (elution:cyclohexane/ethyl acetate 95/5).

Compound 331-[4-hexyloxyphenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one

This compound was synthesized from1-[4-hexyloxyphenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one(compound 32) according to general method 5 described earlier.

Purification was by recrystallization in methanol.

1H NMR DMSO δ ppm: 0.88 (t, J=6.33 Hz, 3H), 1.30 (m, 4H), 1.39 (s, 6H),1.44 (m, 2H), 1.73 (m, 2H), 2.22 (s, 6H), 4.06 (t, J=6.30 Hz, 2H), 7.06(d, J=8.61 Hz, 2H), 7.56 (s, 2H), 7.58 (d, J=15.5 Hz, 1H), 7.82 (d,J=15.5 Hz, 1H), 8.13 (d, J=6.61 Hz, 2H)

MS (ES-MS): 437.2(M−1)

Compound 342-(3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl)-7-chloro-4H-1-benzopyran-4-one

This compound was synthesized from2-(3,5-dimethyl-4-hydroxyphenyl)-7-chloro-4H-1-benzopyran-4-one(intermediate compound 6) and tertbutyl bromoisobutyrate according togeneral method 4 described earlier.

Purification was by precipitation in the solvent mixturedichloromethane/heptane.

Compound 35 2-(3,5-dimethyl-4-carboxydimethylmethyloxyphenyl)-7-chloro-4H-1-benzopyran-4-one

This compound was synthesized from2-(3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl)-7-chloro-4H-1-benzopyran-4-one(compound 34) according to general method 5 described earlier.

Purification was by preparative HPLC (reverse phase RP18, Licrospher12μm, elution: water/methanol/trifluoroacetic acid: 22/78/0.1).

1H NMR DMSO δ ppm: 1.24 (s, 6H), 2.28 (s, 6H), 7.02 (s, 1H), 7.56 (dd,J=8.71 Hz, J=1.75 Hz, 1H), 7.85 (s, 2H), 8.03 (d, J=1.75 Hz, 1H), 8.06(d, J=8.71 Hz, 1H)

MS (Maldi-Tof): 387.1(M+1)

Compound 361-[2-methyloxy-4-chlorophenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one

This compound was synthesized from1-[2-methyloxy-4-chlorophenyl]-3-[3,5-dimethyl-4-hydroxydimethylmethyloxyphenyl]prop-2-en-1-one(intermediate compound 7) and tertbutyl bromoisobutyrate according togeneral method 4 described earlier.

Purification was by chromatography on silica gel (elution:cyclohexane/ethyl acetate 9:1).

Compound 371-[2-methyloxy-4-chlorophenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one

This compound was synthesized from1-[2-methoxy-4-chlorophenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one(compound 36) according to general method 5 described earlier.

Purification was by chromatography on silica gel (elution:dichloromethane/methanol 98:2)

1H NMR DMSO δ ppm: 1.38 (s, 6H), 2.19 (s, 6H), 3.89 (s, 3H), 7.12 (dd,J=7.98 Hz, J=1.71 Hz, 1H), 7.23 (d, J=15.56 Hz, 1H), 7.29 (s, J=1.71 Hz,1H), 7.38 (d, J=15.7 Hz, 1H), 7.41 (s, 2H), 7.48 (d, J=7.98 Hz, 1H)

MS (ES-SM): 401.2(M−1)

Compound 381-[4-heptylphenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one

This compound was synthesized from1-[4-heptylphenyl]-3-[3,5-dimethyl-4-hydroxyphenyl]prop-2-en-1-one(intermediate compound 9) and tertbutyl bromoisobutyrate according togeneral method 4 described earlier.

Purification was by chromatography on silica gel (elution:cyclohexane/ethyl acetate 9/1)

Compound 391-[4-heptylphenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one

This compound was synthesized from1-[4-heptylphenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one(compound 38) and tertbutyl bromoisobutyrate according to general method4 described earlier.

Purification was by chromatography on silica gel (elution:dichloromethane/methanol 98/2)

1H NMR DMSO δ ppm: 0.85 (m, 3H), 1.30-1.24 (m, 8H), 1.39 (s, 6H), 1.60(m, 2H), 2.22 (s, 6H), 2.67 (t, 2H, J=7.4 Hz), 7.37 (d, J=8.04 Hz, 2H),7.57 (s, 2H), 7.62 (d, J=15.66 Hz, 1H), 7.82 (d, J=15.69 Hz, 1H), 8.07(d, J=8.07 Hz, 2H)

MS (ES-MS): 435.3(M−1)

Compound 401-[4-bromophenyl]-3-[3,5-dimethyl]-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one

This compound was synthesized from1-[4-bromophenyl]-3-[3,5-dimethyl-4-hydroxyphenyl]prop-2-en-1-one(intermediate compound 8) and tertbutyl bromoisobutyrate according togeneral method 4 described earlier.

Purification was by chromatography on silica gel (elution:cyclohexane/ethyl acetate 9/1)

Compound 41 1-[4-bromophenyl -3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one

This compound was synthesized from1-[4-bromophenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one(compound 40) according to general method 5 described earlier.

Purification was by chromatography on silica gel (elution:dichloromethane/methanol 98:2)

1H NMR DMSO δ ppm: 1.39 (s, 6H), 2.22 (s, 6H), 7.58 (s, 2H), 7.65 (d,J=15.39 Hz, 1H), 7.84-7.77 (m, 3H), 8.09 (d, J=8.19 Hz, 1H), 13.01 (s,1H)

MS (ES-MS): 417.2 (M−1)

Compound 421-[2-hydroxyphenyl-3-[3,5-dimethyl-4-isopropyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one

1-[2-hydroxyphenyl]-3-[4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one(compound 4; 1 eq) was dissolved in dichloromethane.Dichloromethylmethyl ether (3 eq) was added and the mixture was keptunder reflux for 8 hours. The solvent and excess reagent were eliminatedby vacuum evaporation. The evaporation residue was taken up inisopropanol (50 eq) stirred for 12 hours at room temperature and theisopropanol was then eliminated by vacuum evaporation.

Purification was by chromatography on silica gel (elution: toluene/ethylacetate 7:3)

1H NMR CDCl₃ δppm: 1.21 (d, J=6.09 Hz, 6H), 1.65 (s, 6H), 5.10 (sept,J=6.10 Hz, 1H), 6.86 (d, J=8.65 Hz, 2H), 6.95 (m, 1H), 7.02 (dd, J=8.65Hz, J=1.53 Hz, 1H), 7.48 (m, 1H), 7.54 (d, J=15.25 Hz, 1H), 7.57 (d,J=8.65 Hz, 2H), 7.87 (d, J=15.25 Hz, 1H), 7.93 (d, J=8.40 Hz, 1H), 12.94(signal exchangeable D₂O, 1H)

MS (Maldi-Tof): 369.1(M+1)

Example b 2 Evaluation of PPAR Activation In Vitro

The inventive compounds which were tested are the compounds whosepreparation is described in the above examples.

Nuclear receptors of the PPAR subfamily which are activated by two majorclasses of pharmaceuticals—fibrates and glitazones, widely used in theclinic for the treatment of dyslipidemias and diabetes—play an importantrole in lipid and glucose homeostasis. The following experimental datashow that the inventive compounds activate PPARα and PPARγ in vitro.

PPAR activation was tested in vitro in RK13 fibroblast cell lines bymeasuring the transcriptional activity of chimeras composed of the DNAbinding domain of the yeast gal4 transcription factor and the ligandbinding domain of the different PPARs. These latter results were thenconfirmed in cell lines according to the following protocols:

The example is given for RK13 cells.

a. Culture Protocols

RK13 cells were from ECACC (Porton Down, UK) and were grown in DMEMmedium supplemented with 10% (VN) fetal calf serum, 100 U/mI penicillin(Gibco, Paisley, UK) and 2 mM L-glutamine (Gibco, Paisley, UK). Theculture medium was changed every two days. Cells were kept at 37° C. ina humidified 95% air/5% CO₂ atmosphere.

b. Description of Plasmids Used for Transfection

The plasmids pG5TkpGL3, pRL-CMV, pGal4-hPPARα, pGal4-hPPARγ and pGal4-φhave been described by Raspe, Madsen et al. (1999). The pGal4-mPPARα andpGal4-hPPARγ constructs were obtained by cloning into the pGal4-φ vectorof PCR-amplified DNA fragments corresponding to the DEF domains of thehuman PPARα and PPARγ nuclear receptors.

c. Transfection

RK13 cells were seeded in 24-well culture dishes at 5×10⁴ cells/well andtransfected for 2 hours with the reporter plasmid pG5TkpGL3 (50ng/well), the expression vectors pGal4-φ, pGal4-mPPARα, pGal4-hPPARα,pGal4-hPPARγ (100 ng/well) and the transfection efficiency controlvector pRL-CMV (1 ng/well) according to the previously describedprotocol (Raspe, Madsen et al. 1999), then incubated for 36 hours withthe test compounds. At the end of the experiment, the cells were lysed(Gibco, Paisley, UK) and luciferase activity was determined with aDual-Luciferase™ Reporter Assay System kit (Promega, Madison, Wis., USA)according to the supplier's instructions as previously described (Raspe,Madsen et al. 1999).

The inventors demonstrate an increase in luciferase activity in cellstreated with the inventive compounds and transfected with thepGal4-hPPARα plasmid. Said induction of luciferase activity indicatesthat the inventive compounds are activators of PPARα.

The results are exemplified in FIGS. 2-1, 2-2, 2-3, 2-4, 2-5, 2-6 whichillustrate the PPARα activator properties of inventive compounds 3, 4,7, 8, 9, 11, 12, 13, 14, 17, 19, 20, 21, 22, 23, 24, 25, 26, 29, 31, 33,37, 38, 41.

The inventors demonstrate an increase in luciferase activity in cellstreated with the inventive compounds and transfected with thepGal4-hPPARγ plasmid. Said induction of luciferase activity indicatesthat the inventive compounds are activators of PPARγ.

The results are exemplified in FIG. 2-7 which illustrates the PPARγactivator properties of inventive compounds 17, 33 and 29.

One aspect of the invention is illustrated by the treatment of diseaseslike atherosclerosis and psoriasis the manifestations of which arevascular and cutaneous, respectively. These two pathologies arecharacterized by chronic systemic inflammation and uncontrolled cellproliferation (smooth muscle cells in the case of atherosclerosis andepidermal keratinocytes in psoriasis). These two pathologies have incommon the expression of inflammatory cytokines, mediated by atranscription factor of the inflammatory response NF-κB, AP-1 and NFAT(Komuves, Hanley et al. 2000; Neve, Fruchart et al. 2000). Bydown-regulating the NF-κB and AP-1 signalling pathway, PPARα inhibitsthe expression of genes involved in the inflammatory response such asthe genes coding for interleukin-6, cyclooxygenase-2 and endothelin-1and therefore impedes the mobilization of monocytes and spumous cells tothe atheromatous lesions.

Example 3 Evaluation of the Effects on Lipid Metabolism In Vivo

The inventive compounds which were tested are the compounds whosepreparation is described in the above examples.

Fibrates, widely used in the clinic for the treatment of dyslipidemiasunderlying the development of atherosclerosis, one of the leading causesof morbidity and mortality in the industrialized world, are potentactivators of the PPARα nuclear receptor, which regulates the expressionof genes involved in lipid transport (apolipoproteins such as Apo AI,Apo AII and Apo CIII, membrane transporters such as FAT) and catabolism(ACO, CPT-I and CPT-II). In humans and rodents, treatment with PPARαactivators therefore leads to a decrease in circulating levels ofcholesterol and triglycerides.

The following protocols were designed to demonstrate a decrease incirculating triglyceride and cholesterol levels as well as the interestof the inventive compounds in a context of preventing and/or treatingcardiovascular diseases.

a) Treatment of Animals

Apo E2/E2 transgenic mice were kept on a 12-hour light/dark cycle at aconstant temperature of 20±3° C. After a 1 week acclimation, the micewere weighed and divided into groups of 6 animals selected so that bodyweight would be uniformly distributed. The test compounds were suspendedin carboxymethylcellulose and administered by intragastric gavage at theindicated doses, once a day for 7 or 8 days. Animals had access to foodand water ad libitum. At the end of the experiment the animals wereweighed and sacrificed under anesthesia. Blood was collected on EDTA.Plasma was prepared by centrifugation at 3000 rpm for 20 minutes. Liversamples were taken and stored frozen in liquid nitrogen for subsequentanalysis.

b) Measurement of Serum Lipids and Apolipoproteins

Serum lipid concentrations (total cholesterol and free cholesterol,triglycerides and phospholipids) were determined by a colorimetric assay(Boehringer, Mannheim, Germany) according to the supplier'sinstructions. Serum concentrations of apolipoproteins AI, AII and CIIIwere determined as previously described (Raspe et al. J. Lipid Res. 40,2099-2110, 1999, Asset G et al., Lipids, 34, 39-44, 1999).

The results are exemplified in FIGS. 3-1, 3-2, 3-3, 3-4, 3-5, 3-6, 3-7,3-8, 3-9, 3-10, 3-11 et 3-12 which illustrate the activity of inventivecompounds 7, 17, 29, 33 and 41 on triglyceride and cholesterolmetabolism.

c) RNA Analysis

Total RNA was isolated from liver specimens by extraction with a mixtureof guanidine thiocyanate/acid phenol/chloroform according to thepreviously described protocol (Raspe et al. J. Lipid Res. 40, 2099-2110,1999). Messenger RNA was quantified by quantitative RT-PCR with a LightCycler Fast Start DNA Master Sybr Green I kit (Hoffman-La Roche, Basel,Switzerland) on a Light Cycler System (Hoffman-La Roche, Basel,Switzerland). Primer pairs specific for the genes ACO, Apo CIII and ApoAII were used as probes. Primer pairs specific for the genes 36B4,β-actin and cyclophilin were used as control probes. Alternatively,total RNA was analyzed by Northern Blot or Dot Blot according to thepreviously described protocol (Raspe et al. J. Lipid Res. 40, 2099-2110,1999).

Example 4 Evaluation of the Antioxidant Properties of the InventiveCompounds

A particularly advantageous aspect of the invention is illustrated bythe role of the intrinsic antioxidant properties of the compounds usedin the inventive compositions in the control of oxidative stress. Thisoriginal association between the PPARα agonist property and theantioxidant property represents an effective means of treatingpathologies related to an alteration of the redox status of a cell. Thisillustration applies in particular to a pathology such as Alzheimer'sdisease in which free radicals play a decisive role.

In patients with Alzheimer's disease, oxidative status is modified inbrain cells. Free radicals thus cause lipid peroxidation as well asoxidation of proteins and nucleic acids (DNA/RNA). Said oxidations alterthe biological properties of biomolecules and lead to neuronaldegeneration (Butterfield, Drake et al. 2001). NF-κB is a transcriptionfactor known to be sensitive to the redox status of cells.

Therefore it is closely involved in the response to oxidative stressbecause it allows activation of the target genes of inflammation(Butterfield, Drake et al. 2001). The compounds used in the inventivecompositions therefore have the original property of preventingactivation of the NF-κB pathway at two different levels, by inhibitingits activation by free radicals (antioxidant) but also by preventing itstranscriptional activity (PPARα agonist).

The inventive compounds represent a novel means of fighting against theeffects of ageing and more particularly against the effects ofUV-induced photo-ageing where free radicals actively participate in thepathogenesis of disorders ranging from skin erythema and wrinkleformation to more serious pathologies like skin cancer (basal cell andsquamous cell carcinoma as well as melanoma).

Metabolism is what underlies the production of free radicals, butenvironmental factors like excitatory ionizing radiation (ultraviolet)or inflammatory mediators (cytokines), chemotherapeutic drugs, andhyperthermia are potent activators of free radical species and produce adisequilibrium in the redox balance of the cell. When the stress issevere, survival of the cell depends on its capacity to adapt, resistthe stress and degrade damaged molecules. During ageing, the capacity ofcells to defend themselves appropriately against an oxidative attack iscrucial, and so an increase in cells' capacity to resist such attacksshould help provide a solution to fight against the occurrence of theeffects of ageing and should promote an increase in the longevity of theorganism.

Solar radiation can alter the composition of certain molecules in thebody. UVB was long considered to be the sole cause of the sun'sdeleterious effects on the body. It is now known that UVA radiation canhave a direct adverse effect but above all that it potentiates theeffects of UVB. The principal molecules vulnerable to alteration, oftenin a harmful way but also in a beneficial way, are:

-   -   DNA, in which thymine dimers can form under the action of UVB.        Although DNA does not absorb UVA rays, the latter can damage the        genetic material and therefore be mutagenic. A pathology such as        Xeroderma pigmentosum, which is due to an absence or an        alteration of DNA repair mechanisms, predisposes to the        development of cancer of basal keratinocytes.    -   Proteins, the spatial conformation of which can be altered. Many        proteins can be inactivated in this manner: enzymes,        transporters, ion channels, cytoskeletal proteins, receptors.        Said alterations can be induced by UVA and UVB radiation.    -   Lipids, which can undergo UVA-induced peroxidation, said        peroxidation being proportional to the degree of fatty acid        unsaturation.

The following protocols were designed to demonstrate the intrinsicantioxidant properties of the compounds used in the inventivecompositions for prevention and/or treatment of disorders related tooxidative stress.

1. Protection Against LDL Oxidation by Copper:

The inventive compounds which were tested are the compounds whosepreparation is described in the above examples.

LDL oxidation is an important alteration and plays a predominant role inthe establishment and development of atherosclerosis (Jurgens, Hoff etal. 1987). The following protocol allows to demonstrate the antioxidantproperties of compounds. Unless otherwise indicated, the reagents werefrom Sigma (St Quentin, France).

LDL were prepared according to the method described by Lebeau et al.(Lebeau, Furman et al. 2000).

The solutions of test compounds were prepared at 10⁻² M concentration inbicarbonate buffer (pH 9) and diluted in PBS to obtain finalconcentrations ranging from 0.1 to 100 μM for a total ethanolconcentration of 1% (V/V).

Prior to oxidation, EDTA was removed from the LDL preparation bydialysis. Oxidation then took place at 30° C. by addition of 100 μl of16.6 μM CuSO₄ solution to 160 μL of LDL (125 μg protein/ml) and 20 μl ofa test compound solution. The formation of dienes, the species underobservation, was followed by measuring optical density at 234 nm in thesamples treated with the compounds but in the presence or absence ofcopper. Optical density at 234 nm was measured every 10 minutes for 8hours in a thermostated spectrophotometer (Tecan Ultra 380). Theanalyses were performed in triplicate. The compounds were considered tohave antioxidant activity when they induced a longer lag phase andreduced the rate of oxidation and the amount of dienes formed incomparison with the control sample. The inventors demonstrate that theinventive compounds have at least one of the above-described antioxidantproperties indicating that the inventive compounds have intrinsicantioxidant activity. FIGS. 1, 2 and 3 give an example of the resultsillustrating the antioxidant properties of compounds 2 and 5.

Results are given in FIGS. 1-1, 1-2, 1-3, 1-4, 1-5, 1-6, 1-7, 1-8, 1-9,1-10, 1-11, 1-12, 1-13 and 1-14 illustrating the antioxidant propertiesof inventive compounds 2, 3, 4, 5, 6, 7, 8, 9, 10, 14, 17, 18, 19, 21,22, 25, 29, 31, 33, 35, 37, 38 and 41.

2. Evaluation of Protection Conferred by the Inventive Compounds againstLipid Peroxidation:

The inventive compounds which were tested are the compounds whosepreparation is described in the above examples.

LDL oxidation was determined by the TBARS method.

According to the same principle described earlier, LDL were oxidizedwith CuSO₄ and lipid peroxidation was determined as follows:

TBARS were measured by a spectrophotometric method, lipidhydroperoxidation was measured using lipid-dependent peroxidation ofiodide to iodine. The results are expressed as nmol of malondialdehyde(MDA) or as nmol of hydroperoxide/mg of protein.

The previous results obtained by measuring inhibition of conjugateddiene formation were confirmed by the experiments measuring LDL lipidperoxidation.

The inventive compounds also effectively protected LDL against lipidperoxidation induced by copper (oxidizing agent).

BIBLIOGRAPHY

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1-12. (canceled)
 13. A method for the treatment of a pathology relatedto inflammation, neurodegeneration, cell proliferation and/ordifferentiation and/or skin or central nervous system ageing, comprisingadministering to a subject in need thereof an effective amount of apharmaceutical composition comprising at least one substituted1,3-diphenylprop-2-en-1-one derivative represented by formula (I) below:

wherein: X1 represents a halogen or a —R1 group or a group correspondingto the following formula: -G1-R1, X2 represents a hydrogen atom or ahydroxy group or an unsubstituted alkyloxy group or a thiol group or analkylthio group, X3 represents a —R3 group, X4 represents a groupcorresponding to the following formula: -G4-R4, X5 represents a —R5group, X6 is an oxygen atom, R3 and R5, which are the same or different,represent an unsubstituted alkyl group, R1 represents a hydrogen atom oran alkyl group substituted or not by a substituent having the formula—COOR6, with R6 representing a hydrogen atom or an alkyl group, R4represents a hydrogen atom or an alkyl group substituted or not by asubstituent having the formula —COOR6, with R6 representing a hydrogenatom or an alkyl group, G1 and G4, which are the same or different,represent an oxygen or sulfur atom, with one of the groups R1 and R4represents an alkyl group substituted by a substituent having theformula —COOR6, said alkyl group being bound directly to the ring orbeing associated with a group G according to the formula -GR, theoptical and geometrical isomers, racemates, tautomers, salts, andmixtures thereof.
 14. The method according to claim 13, wherein thederivative corresponds to the cis or trans conformation or a mixturethereof.
 15. The method according to claim 13, wherein X2 is a hydroxygroup or an alkyloxy.
 16. The method according to claim 13, wherein X4is a group corresponding to the formula -G4-R4 and X2 is a hydroxy groupor an alkyloxy group and, G4 and R4 being such as defined in claim 13.17. The method according to claim 13, wherein X1 represents a —R1 groupor a group corresponding to the formula -G1-R1, where R1 is an alkylgroup substituted by a substituent having the formula —COOR6, and G1 isas defined in claim
 13. 18. The method according to claim 13, wherein X1is a -G1-R1 group.
 19. The method according to claim 13, wherein X1 is a-G1-R1 group in which G1 is an oxygen atom.
 20. The method according toclaim 13, wherein X4 represents a group corresponding to the formula-G4-R4, where R4 is an alkyl group substituted by a substituent havingthe formula —COOR6, and G4 being such as defined in claim
 13. 21. Themethod according to claim 13, wherein X4 is a -G4-R4 group in which G4is an oxygen atom.
 22. The method according to claim 13, wherein X4 is a-G4-R4 group in which G4 is an oxygen atom, and X3 or X5 respectivelyrepresents R3 and R5, with R3 and R5 being an alkyl group having fromone to seven carbon atoms.
 23. The method according to claim 13, whereinX1 represents a halogen.
 24. The method according to claim 13, whereinX1 represents a —R1 group with R1 being a C1 to C4 alkyl groupsubstituted or not by at least one substituent having the formula—COOR6.
 25. The method according to claim 13, wherein X1 represents a-G1R1 group with R1 being a C1 to C3 alkyl group substituted or not byat least one substituent having the formula —COOR6.
 26. The methodaccording to claim 13, wherein X1 represents a —R1 group with R1 beingan unsubstituted C5 to C24 alkyl group.
 27. The method according toclaim 13, wherein X1 represents a -G1R1 group with R1 being anunsubstituted C4 to C24 alkyl group.
 28. The method according to claim13, wherein X1 or X4 represents OC(CH3)2COOR6.
 29. The method accordingto claim 13, wherein X1 or X4 represents SC(CH3)2COOR6.
 30. The methodaccording to claim 13, wherein the derivative is selected in the groupconsisting of:1-[2-hydroxy-4-carboxydimethylmethyloxyphenyl]-3-[3,5-ditertbutyl-4-hydroxyphenyl]prop-2-en-1-one,1-[2-hydroxy-4-ethyloxycarbonyldimethylmethyloxyphenyl]-3-[3,5-ditertbutyl-4-hydroxyphenyl]prop-2-en-1-one,1-[2-hydroxy-4-carboxydimethylmethyloxyphenyl]-3-[3,5-dimethyl-4-hydroxyphenyl]prop-2-en-1-one,1-[2-hydroxy-4-isopropyloxycarbonyldimethylmethyloxyphenyl]-3-[3,5-dimethyl-4-hydroxyphenyl]prop-2-en-1-one,1-[2-hydroxy-4-chlorophenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one,1-[2-hydroxy-4-chlorophenyl]-3-[3,5-dimethyl-4-isopropyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one,1-[2-hydroxyphenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one,1-[2-hydroxyphenyl]-3-[3,5-dimethyl-4-isopropyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one,1-[2-hydroxy-4-ethoxycarbonyldimethylmethyloxyphenyl]-3-[3,5-ditertbutyl-4-hydroxyphenyl]prop-2-en-1-one,1-[4-chlorophenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one,1-[4-chlorophenyl]-3-[3,5-dimethyl-4-isopropyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one,1-[4-chlorophenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one,1-[4-carboxydimethylmethyloxyphenyl]-3-[3,5-dimethyl-4-hydroxyphenyl]prop-2-en-1-one,1-[2-hydroxy-4-bromophenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one,1-[4-methylthiophenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one,1-[4-methylthiophenyl]-3-[3,5-dimethyl-4-isopropyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one,1-[4-methylthiophenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one,1-[2-methoxyphenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one,1-[2-methoxyphenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one,1-[4-hexyloxyphenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one,1-[4-hexyloxyphenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one,1-[2-methyloxy-4-chlorophenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one,1-[2-methyloxy-4-chlorophenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one,1-[4-heptylphenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one,1-[4-heptylphenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one,1-[4-bromophenyl]-3-[3,5-dimethyl-4-tertbutyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one,1-[4-bromophenyl]-3-[3,5-dimethyl-4-carboxydimethylmethyloxyphenyl]prop-2-en-1-one,1-[2-hydroxyphenyl]-3-[3,5-dimethyl-4-isopropyloxycarbonyldimethylmethyloxyphenyl]prop-2-en-1-one.31. The method according to claim 13, wherein the pathology related toinflammation is selected in the group consisting of atherosclerosis,allergy, asthma, eczema, psoriasis and pruritus.
 32. The methodaccording to claim 13, wherein the pathology related toneurodegeneration is Alzheimer's disease or Parkinson's disease.
 33. Themethod according to claim 13, wherein the pathology related to cellproliferation and/or differentiation is selected in the group consistingof carcinogenesis, psoriasis and atherosclerosis.